EP2313094A1 - New pyridine derivatives as leptin receptor modulator mimetics - Google Patents

New pyridine derivatives as leptin receptor modulator mimetics

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Publication number
EP2313094A1
EP2313094A1 EP09757597A EP09757597A EP2313094A1 EP 2313094 A1 EP2313094 A1 EP 2313094A1 EP 09757597 A EP09757597 A EP 09757597A EP 09757597 A EP09757597 A EP 09757597A EP 2313094 A1 EP2313094 A1 EP 2313094A1
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EP
European Patent Office
Prior art keywords
pyridin
ylmethyl
carbamate
alkyl
hydroxy
Prior art date
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Application number
EP09757597A
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German (de)
French (fr)
Inventor
Michael Higginbottom
Iain Simpson
James Horton
Charles Tyzack
Anne Viet-Anh Horgan
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AstraZeneca AB
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AstraZeneca AB
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Publication of EP2313094A1 publication Critical patent/EP2313094A1/en
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    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
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    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
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    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present application relates to new pyridine derivatives, to pharmaceutical compositions comprising these compounds and to the use of these compounds as leptin receptor modulator mimetics in the preparation of medicaments against conditions associated with weight gain, type 2 diabetes and dyslipidemias.
  • the first line of treatment is to offer diet and life style advice to patients, such as reducing the fat content of their diet and increasing their physical activity.
  • patients may also need to undergo drug therapy to maintain the beneficial results obtained from adapting the aforementioned diet and lifestyle changes.
  • Leptin is a hormone synthesized in fat cells that is believed to act in the hypothalamus to reduce food intake and body weight (see, e.g., Bryson, J. M. (2000) Diabetes, Obesity and Metabolism 2: 83-89).
  • Leptin can also act to increase inflammation by enhancing the release of pro -inflammatory cytokines TNF and IL-6 from inflammatory cells (Zarkesh-Esfahani, H. et al. (2001) J. Immunol. 167: 4593-4599). These agents in turn can contribute to the insulin resistance commonly seen in obese patients by reducing the efficacy of insulin receptor signaling (Lyon, C. J. et al. (2003) Endocrinol. 44: 2195-2200). Continuous low grade inflammation is believed to be associated with obesity (in the presence and absence of insulin resistance and Type II diabetes) (Browning et al.
  • Leptin has also been shown to promote the formation of new blood vessels (angiogenesis) a process implicated in the growth of adipose tissue (Bouloumie A, et al. (1998) Circ. Res. - -
  • Angiogenesis has also been implicated in diabetic retinopathy (Suganami, E. et al. (2004) Diabetes. 53: 2443-2448).
  • Angiogenesis is also believed to be involved with the growth of new blood vessels that feed abnormal tumour cells. Elevated leptin levels have been associated with a number of cancers, in particular breast, prostate and gastrointestinal cancers in humans (Somasundar P. et al. (2004) J. Surg. Res. 116: 337-349).
  • Leptin receptor agonists may also be used in the manufacture of a medicament to promote wound healing (Gorden, P. and Gavrilova, O. (2003) Current Opinion in Pharmacology 3: 655-659). Further, it has been shown that elevating leptin signaling in the brain may represent an approach for the treatment of depressive disorders (Lu, Xin-Yun et al. (2006) PNAS 103: 1593-1598).
  • compounds with leptin receptor agonistic like properties can be useful for the treatment of disorders relating to leptin signaling, as well as conditions associated with weight gain, such as obesity.
  • compounds with leptin receptor antagonistic like properties could be useful for the treatment of inflammation, atherosclerosis, diabetic retinopathy and nephropathy. - -
  • the disclosure relates to a compound of formula (I),
  • each R 1 is independently selected from Ci_ 4 -alkyl, Ci_ 4 -alkoxy, halogen, cyano and CF 3 ;
  • R 2 is Ci_6-alkyl (optionally substituted with one or more substituents selected from hydroxy, halogen and cyano) or -[C(R 4A )(R 4B )] m -R 5 ;
  • R 3 is hydrogen, Ci_ 4 -alkyl or fluoro-Ci_ 4 -alkyl
  • R 4A and R 4B are each independently selected from hydrogen, halogen, hydroxy, Ci_ 4 -alkyl, fluoro-Ci_ 4 -alkyl and hydroxy-Ci_ 4 -alkyl;
  • R 5 is C 3 - 8 -cycloalkyl, C ⁇ -io-aryl, heterocyclyl or heteroaryl, each of which is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro,
  • n 0, 1, 2, 3 or 4;
  • R 2 is Ci_ 6 -alkyl which is substituted with one or more substituents selected from hydroxy, halogen and cyano, more preferably hydroxy.
  • R 2 is 2-hydroxyethyl, l-(hydroxymethyl)-2-methylpropyl or 1 -(hydroxymethyl)-3 -methylbutyl.
  • R 2 is -[C(R 4A )(R 4B )] m -R 5 , wherein R 5 is phenyl, C 3 - 6 - cycloalkyl, tetrahydrofuranyl, indanyl or isoxazolyl, and is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro, CF 3 or Ci_4-alkyl.
  • R 5 is phenyl, C 3 - 6 - cycloalkyl, tetrahydrofuranyl, indanyl or isoxazolyl, and is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro, CF 3 or Ci_4-alkyl.
  • R 5 is cyclopropyl, cyclopentyl, 1-hydroxycyclohexyl, 2-tetra- hydrofuranyl, 3-tetrahydrofuranyl, 2-hydroxy-lH-inden-l-yl or 3,5-dimethylisoxazol-4-yl.
  • n is preferably 0 or 1.
  • R 4A and R 4B are preferably independently selected from hydrogen, methyl and hydroxymethyl.
  • R 3 is preferably hydrogen or methyl.
  • Another aspect of the present disclosure is a compound of formula (I) for use in therapy. - -
  • the invention relates to a compound of formula (I) for use in the treatment or prevention of any of the disorders or conditions described herein.
  • the invention relates to the use of the compounds of formula (I) in the manufacture of a medicament for the treatment or prevention of any of the disorders or conditions described herein.
  • said compounds may be used for the treatment or prevention of a condition that is prevented, treated, or ameliorated by selective action via the leptin receptor.
  • compounds of formula (I) may be used for the treatment or prevention of conditions (in particular, metabolic conditions) that are associated with weight gain.
  • Conditions associated with weight gain include diseases, disorders, or other conditions that have an increased incidence in obese or overweight subjects. Examples include: lipodystrophy, HIV lipodystrophy, diabetes (type 2), insulin resistance, metabolic syndrome, hyperglycemia, hyperinsulinemia, dyslipidemia, hepatic steatosis, hyperphagia, hypertension, hypertriglyceridemia, infertility, a skin disorder associated with weight gain, macular degeneration.
  • compounds of the invention may also be used in the manufacture of a medicament for maintaining weight loss of a subject.
  • compounds of formula (I) which are leptin receptor agonist mimetics may also be used to promote wound healing.
  • compounds of formula (I) which are leptin receptor agonist mimetics may also be used for the treatment or prevention of conditions that cause a decrease in circulating leptin concentrations, and the consequent malfunction of the immune and reproductive systems. Examples of such conditions and malfunctions include severe weight loss, dysmenorrhea, amenorrhea, female infertility, immunodeficiency and conditions associated with low testosterone levels. - -
  • compounds of formula (I) which are leptin receptor agonist mimetics may also be used for the treatment or prevention of conditions caused as a result of leptin deficiency, or a leptin or leptin receptor mutation.
  • compounds of formula (I) which are leptin receptor antagonist mimetics may be used for the treatment or prevention of inflammatory conditions or diseases, low level inflammation associated with obesity and excess plasma leptin and in reducing other complications associated with obesity including atherosclerosis, and for the correction of insulin resistance seen in Metabolic Syndrome and diabetes.
  • compounds of formula (I) which are leptin receptor antagonist mimetics can be used for the treatment or prevention of inflammation caused by or associated with: cancer (such as leukemias, lymphomas, carcinomas, colon cancer, breast cancer, lung cancer, pancreatic cancer, hepatocellular carcinoma, kidney cancer, melanoma, hepatic, lung, breast, and prostate metastases, etc.); auto-immune disease (such as organ transplant rejection, lupus erythematosus, graft v.
  • cancer such as leukemias, lymphomas, carcinomas, colon cancer, breast cancer, lung cancer, pancreatic cancer, hepatocellular carcinoma, kidney cancer, melanoma, hepatic, lung, breast, and prostate metastases, etc.
  • auto-immune disease such as organ transplant rejection, lupus erythematosus, graft v.
  • autoimmune damage including multiple sclerosis, Guillam Barre Syndrome, myasthenia gravis
  • cardiovascular conditions associated with poor tissue perfusion and inflammation such as atheromas, atherosclerosis, stroke, ischaemia-reperfusion injury, claudication, spinal cord injury, congestive heart failure, vasculitis, haemorrhagic shock, vasospasm following subarachnoid haemorrhage, vasospasm following cerebrovascular accident, pleuritis, pericarditis, the cardiovascular complications of diabetes); ischaemia- reperfusion injury, ischaemia and associated inflammation, restenosis following angioplasty and inflammatory aneurysms; epilepsy, neurodegeneration (including Alzheimer's Disease), arthritis (such as rheumatoid arthritis, osteoarthritis
  • bronchoconstriction pulmonary vasoconstriction, impeded respiration, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcosis, cystic fibrosis, pulmonary hypertension, pulmonary vasoconstriction, emphysema, bronchial allergy and/or inflammation, asthma, hay fever, rhinitis, vernal conjunctivitis and adult respiratory distress syndrome); conditions associated with inflammation of the skin (including psoriasis, eczema, ulcers, contact dermatitis); conditions associated with inflammation of the bowel (including Crohn's disease, ulcerative colitis and pyresis, irritable bowel syndrome, inflammatory bowel disease); HIV (particularly HIV infection), cerebral malaria, bacterial meningitis, osteoporosis and other bone resorption diseases, osteoarthritis, infertility from endometriosis, fever and myalgia due to infection, and other conditions mediated by excessive anti-inflammatory cell (including neutr
  • compounds of formula (I) which are leptin receptor antagonists mimetics may be used for the treatment or prevention of macro or micro vascular complications of type 1 or 2 diabetes, retinopathy, nephropathy, autonomic neuropathy, or blood vessel damage caused by ischaemia or atherosclerosis.
  • compounds of formula (I) which are leptin receptor antagonist mimetics may be used to inhibit angiogenesis.
  • Compounds that inhibit angiogenesis may be used for the treatment or prevention of obesity or complications associated with obesity.
  • Compounds that inhibit angiogenesis may be used for the treatment or prevention of complications associated with inflammation diabetic retinopathy, or tumour growth particularly in breast, prostate or gastrointestinal cancer.
  • the disclosure relates to a method for the treatment or prevention of any of the disorders or conditions described herein, which includes administering to a subject (e.g., a subject in need thereof, e.g., a mammal) an effective amount of a compound of formula I.
  • a subject e.g., a subject in need thereof, e.g., a mammal
  • Methods delineated herein include those wherein the subject is identified as in need of a particular stated treatment. Identifying a subject in need of such treatment can be in the - -
  • judgment of a subject or a health care professional can be subjective (e.g. opinion) or objective (e.g. measurable by a test or diagnostic method).
  • the methods herein include those further comprising monitoring subject response to the treatment administrations.
  • monitoring may include periodic sampling of subject tissue, fluids, specimens, cells, proteins, chemical markers, genetic materials, etc. as markers or indicators of the treatment regimen.
  • the subject is prescreened or identified as in need of such treatment by assessment for a relevant marker or indicator of suitability for such treatment.
  • the invention provides a method of monitoring treatment progress.
  • the method includes the step of determining a level of diagnostic marker (Marker) (e.g., any target or cell type delineated herein modulated by a compound herein) or diagnostic measurement (e.g., screen, assay) in a subject suffering from or susceptible to a disorder or symptoms thereof delineated herein, in which the subject has been administered a therapeutic amount of a compound herein sufficient to treat the disease or symptoms thereof.
  • a level of diagnostic marker e.g., any target or cell type delineated herein modulated by a compound herein
  • diagnostic measurement e.g., screen, assay
  • a second level of Marker in the subject is determined at a time point later than the determination of the first level, and the two levels are compared to monitor the course of disease or the efficacy of the therapy.
  • a pre-treatment level of Marker in the subject is determined prior to beginning treatment according to this invention; this pre-treatment level of Marker can then be compared to the level of Marker in the subject after the treatment commences, to determine the efficacy of the treatment.
  • a level of Marker or Marker activity in a subject is determined at least once.
  • Comparison of Marker levels may be useful in determining whether therapy according to the disclosure is having the desired effect, and thereby permitting adjustment of dosage levels as appropriate.
  • Determination of Marker levels may be performed using any suitable sampling/expression assay method known in the art or described herein. Preferably, a tissue or fluid sample is first removed from a subject. Examples of suitable samples include blood, urine, tissue, mouth or cheek cells, and hair samples containing roots. Other suitable samples would be known to the person skilled in the art. Determination of protein - -
  • mRNA levels and/or mRNA levels (e.g., Marker levels) in the sample can be performed using any suitable technique known in the art, including, but not limited to, enzyme immunoassay, ELISA, radio labeling/assay techniques, blotting/chemiluminescence methods, real-time PCR, and the like.
  • a compound of formula (I) it may be advantageous if a compound of formula (I) is able to penetrate the central nervous system. In other embodiments, it may be advantageous if a compound of formula (I) is not able to penetrate the CNS.
  • compounds that are leptin receptor agonist mimetics may be particularly useful for the treatment or prevention of obesity, insulin resistance, or diabetes (particularly glucose intolerance) if these compounds can penetrate the CNS.
  • a person of ordinary skill in the art can readily determine whether a compound can penetrate the CNS. A suitable method that may be used is described in the Biological Methods section.
  • a leptin receptor response may be measured in any suitable way. In vitro, this may be done be measuring leptin receptor signaling. For example, phosphorylation of Akt, STAT3, STAT5, MAPK, shp2 or the leptin receptor in response to binding of leptin or a compound of the invention to the leptin receptor may be measured. The extent of phosphorylation of Akt, STAT3, STAT5, MAPK, shp2 or the leptin receptor may be determined for example by Western blotting or by ELISA. Alternatively, a STAT reporter assay may be used, for example STAT driven luciferase expression. A cell line expressing the leptin receptor may be used for such assays. In vivo, leptin receptor response may be measured by determining the reduction in food intake and body weight after administration of leptin or a compound of formula (I).
  • the Biological Methods below describe assays and methods that can be used to determine whether a compound of formula (I) is a leptin receptor agonist mimetic or a leptin receptor antagonist mimetic.
  • a compound of formula (I) may be administered with or without other therapeutic agents.
  • a compound may be administered with an anti-inflammatory agent (for example, disease modifying anti-rheumatic drugs such as methotrexate, sulphasalazine and cytokine inactivating agents, steroids, NSAIDs, - -
  • an anti-inflammatory agent for example, disease modifying anti-rheumatic drugs such as methotrexate, sulphasalazine and cytokine inactivating agents, steroids, NSAIDs, - -
  • a compound may be administered with a cytotoxic agent (for example, methotrexate, cyclophosphamide) or another anti-tumour drug.
  • a cytotoxic agent for example, methotrexate, cyclophosphamide
  • another anti-tumour drug for example, methotrexate, cyclophosphamide
  • Compounds of formula (I) may be radiolabeled (for example with tritium or radioactive iodine) for in vitro or in vivo applications, such as receptor displacement studies or receptor imaging.
  • Ci_6-alkyl denotes a straight or branched alkyl group having from 1 to 4 carbon atoms.
  • examples of said Ci_ 6 -alkyl include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl and t-butyl, as well as straight- and branched-chain pentyl and hexyl.
  • Ci_ 6-alkyl For parts of the range "Ci_6-alkyl" all subgroups thereof are contemplated such as Ci_ 5 -alkyl, Ci_ 4 -alkyl, Ci_ 3 -alkyl, Ci_ 2 -alkyl, C 2 - 6 -alkyl, C 2 - 5 -alkyl, C 2 _ 4 -alkyl, C 2 _ 3 -alkyl, C 3 - 6 -alkyl, C 3 - 5 -alkyl,C 3 - 4 -alkyl, C 4 - 6 -alkyl, C 4 - 5 -alkyl and C 5 . 6 -alkyl.
  • hydroxy-Ci_4-alkyl denotes a straight or branched Ci_4-alkyl group that has a hydrogen atom thereof replaced with OH.
  • examples of said hydroxy-Ci_ 4 -alkyl include hydroxymethyl, 2-hydroxyethyl and 2-hydroxypropyl.
  • fluoro-Ci_4-alkyl denotes a straight or branched Ci_4-alkyl group substituted by one or more fluorine atoms.
  • fluoro-Ci_ 4 -alkyl include fluoromethyl, trifluoromethyl, 2-fluoroethyl and 2,2,2- trifluoroethyl.
  • Ci_4-alkoxy denotes a straight or branched alkoxy group having from 1 to 4 carbon atoms.
  • examples of said Ci_4-alkoxy include methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, iso-butoxy, sec-butoxy and t-butoxy.
  • Ci_4-alkoxy all subgroups thereof are contemplated such as C 1-3 - alkoxy, Ci_2-alkoxy, C2-4-alkoxy, C2- 3 -alkoxy and C 3 _4-alkoxy.
  • C 3 _8-cycloalkyl denotes a mono- or bicyclic saturated hydrocarbon ring system having 3 to 8 carbon atoms. Bicyclic ring systems can be either fused or bridged. In a bridged cycloalkyl ring system, two non- - -
  • C3_8-cycloalkyl examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, as well as bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl and bicyclo[3.2.1]octyl.
  • Cs-s-cycloalkyl For parts of the range "Cs-s-cycloalkyl" all subgroups thereof are contemplated such as C 3 - 7 -cycloalkyl, C 3 _ 6 -cycloalkyl, C 3 _ 5 -cycloalkyl, C 3 _ 4 -cycloalkyl, C 4 _ 8 -cycloalkyl, C 4 _ 7 -cycloalkyl, C 4 _ 6 -cycloalkyl, C 4 _ 5 -cycloalkyl, Cs_ 8 -cycloalkyl, Cs_ 7 -cycloalkyl, Cs_ 6 -cycloalkyl, C 6 - 8 -cycloalkyl and C 6 - 7 -cycloalkyl.
  • C ⁇ -io-aryl refers to a mono- or bicyclic hydrocarbon ring system comprising 6 to 10 ring atoms and wherein at least one ring is an aromatic ring.
  • Examples of C ⁇ -io-aryl include phenyl, indenyl, 2,3-dihydroindenyl (indanyl), 1-naphthyl, 2-naphthyl or 1,2,3,4-tetrahydronaphthyl.
  • heterocyclyl refers to a stable, fully saturated or partially unsaturated mono- or bicyclic ring system having 3 to 8 ring atoms with at least one heteroatom such as O, N, or S, and the remaining ring atoms are carbon.
  • heterocyclyl examples include piperidinyl, tetrahydropyranyl, tetrahydrofuranyl, azepinyl, azetidinyl, pyrrolidinyl, morpholinyl, imidazolinyl, thiomorpholinyl, pyranyl, dioxanyl, piperazinyl, as well as l-azabicyclo[2.2.2]octane, l-azabicyclo[2.2.1]heptane and azabicyclo[2.2.2]oct-2-ene.
  • An exemplary heterocyclic group containing sulfur in oxidized form is thiomorpholine- 1 , 1 -dioxide.
  • heteroaryl refers to a mono- or bicyclic heteroaromatic ring system having 5 to 10 ring atoms in which one or more of the ring atoms are other than carbon, such as nitrogen, sulphur or oxygen. Only one ring need to be aromatic and said heteroaryl moiety can be linked to the remainder of the molecule via a carbon or nitrogen atom in any ring.
  • heteroaryl examples include furyl, pyrrolyl, thienyl, oxazolyl, isoxazolyl, imidazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, tetrazolyl, quinazolinyl, indolyl, isoindolyl, 1,3-dihydro-isoindolyl, pyrazolyl, pyridazinyl, quinolinyl, quinoxalinyl, thiadiazolyl, benzofuranyl, 2,3-dihydrobenzofuranyl, 1,3- benzodioxolyl, 1 ,4-benzodioxinyl, 2,3-dihydro-l,4-benzodioxinyl, benzothiazolyl, benzimidazolyl, benzothiadiazolyl, benzotriazolyl, indolinyl, isoin
  • Niro refers to the -NO 2 radical.
  • Cyano refers to the -CN radical.
  • mammal includes organisms, which include mice, rats, cows, sheep, pigs, rabbits, goats, and horses, monkeys, dogs, cats, and preferably humans.
  • the subject may be a human subject or a non human animal, particularly a domesticated animal, such as a dog.
  • “Pharmaceutically acceptable” means being useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable and includes being useful for veterinary use as well as human pharmaceutical use.
  • Treatment includes prophylaxis of the named disorder or condition, or amelioration or elimination of the disorder once it has been established.
  • “An effective amount” refers to an amount of a compound that confers a therapeutic effect (e.g., treats, controls, ameliorates, prevents, delays the onset of, or reduces the risk of developing a disease, disorder, or condition or symptoms thereof) on the treated subject.
  • the therapeutic effect may be objective (i.e., measurable by some test or marker) or subjective (i.e., subject gives an indication of or feels an effect).
  • Prodrugs refers to compounds that may be converted under physiological conditions or by so lvo lysis to a biologically active compound of formula (I).
  • a prodrug may be inactive when administered to a subject in need thereof, but is converted in vivo to an active compound of formula (I).
  • Prodrugs are typically rapidly transformed in vivo to yield the parent compound, e.g. by hydrolysis in the blood.
  • the prodrug compound usually offers advantages of solubility, tissue compatibility or delayed release in a mammalian organism (see Silverman, R. B., The Organic Chemistry of Drug Design and Drug Action, 2 nd Ed., Elsevier Academic Press (2004), pp. 498-549).
  • Prodrugs may be prepared by modifying functional groups, such as a hydroxy, amino or mercapto groups, present in a compound of formula (I) in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compound.
  • Examples of prodrugs include, but are not limited to, acetate, formate and succinate derivatives of hydroxy functional groups or phenyl carbamate derivatives of amino functional groups.
  • a given chemical formula or name shall also encompass all salts, hydrates, solvates, N-oxides and prodrug forms thereof. Further, a given chemical formula or name shall encompass all tautomeric and stereoisomeric forms thereof.
  • Stereoisomers include enantiomers and diastereomers. Enantiomers can be present in their pure forms, or as racemic (equal) or unequal mixtures of two enantiomers. Diastereomers can be present in their pure forms, or as mixtures of diastereomers. Diastereomers also include geometrical isomers, which can be present in their pure cis or trans forms or as mixtures of those.
  • the compounds of formula (I) may be used as such or, where appropriate, as pharmacologically acceptable salts (acid or base addition salts) thereof.
  • pharmacologically acceptable addition salts mentioned below are meant to comprise the therapeutically active non-toxic acid and base addition salt forms that the compounds are able to form.
  • Compounds that have basic properties can be converted to their pharmaceutically acceptable acid addition salts by treating the base form with an appropriate acid.
  • Exemplary acids include inorganic acids, such as hydrogen chloride, hydrogen bromide, hydrogen iodide, sulphuric acid, phosphoric acid; and organic acids such as formic acid, acetic acid, propanoic acid, hydroxyacetic acid, lactic acid, pyruvic acid, glycolic acid, maleic acid, malonic acid, oxalic acid, benzenesulphonic acid, toluenesulphonic acid, methanesulphonic acid, trifluoroacetic acid, fumaric acid, succinic acid, malic acid, tartaric acid, citric acid, salicylic acid, / ⁇ -aminosalicylic acid, pamoic acid, benzoic acid, ascorbic acid and the like.
  • organic acids such as formic acid, acetic acid, propanoic acid, hydroxyacetic acid, lactic acid, pyruvic acid, glycolic acid, maleic acid, malonic acid, oxalic acid, benzenesulphonic acid, tolu
  • Exemplary base addition salt forms are the sodium, potassium, calcium salts, and salts with pharmaceutically acceptable amines such as, for example, ammonia, alkylamines, benzathine, and amino acids, such as, e.g. arginine and lysine.
  • the term addition salt as used herein also comprises solvates which the compounds and salts thereof are able to form, such as, for example, hydrates, alcoholates and the like.
  • the compounds of formula (I) are formulated into pharmaceutical formulations for various modes of administration. It will be appreciated that the compounds may be administered together with a physiologically acceptable carrier, excipient, or diluent.
  • the pharmaceutical compositions may be administered by any - -
  • suitable route preferably by oral, rectal, nasal, topical (including buccal and sublingual), sublingual, transdermal, intrathecal, transmucosal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration.
  • compositions may conveniently be presented in unit dosage form, e.g., tablets and sustained release capsules, and in liposomes, and may be prepared by any methods well known in the art of pharmacy.
  • Pharmaceutical formulations are usually prepared by mixing the active substance, or a pharmaceutically acceptable salt thereof, with conventional pharmaceutically acceptable carriers, diluents or excipients.
  • excipients are water, gelatin, gum arabicum, lactose, microcrystalline cellulose, starch, sodium starch glycolate, calcium hydrogen phosphate, magnesium stearate, talcum, colloidal silicon dioxide, and the like.
  • Such formulations may also contain other pharmacologically active agents, and conventional additives, such as stabilizers, wetting agents, emulsifiers, flavouring agents, buffers, and the like.
  • the amount of active compounds is between 0.1-95% by weight of the preparation, preferably between 0.2-20% by weight in preparations for parenteral use and more preferably between 1-50% by weight in preparations for oral administration.
  • the formulations can be further prepared by known methods such as granulation, compression, microencapsulation, spray coating, etc.
  • the formulations may be prepared by conventional methods in the dosage form of tablets, capsules, granules, powders, syrups, suspensions, suppositories or injections.
  • Liquid formulations may be prepared by dissolving or suspending the active substance in water or other suitable vehicles. Tablets and granules may be coated in a conventional manner. To maintain therapeutically effective plasma concentrations for extended periods of time, the compounds may be incorporated into slow release formulations.
  • the dose level and frequency of dosage of the specific compound will vary depending on a variety of factors including the potency of the specific compound employed, the metabolic stability and length of action of that compound, the patient's age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the condition to be treated, and the patient undergoing therapy.
  • the daily dosage may, for example, range from about 0.001 mg to about 100 mg per kilo of body weight, administered singly or multiply in doses, e.g. from about 0.01 mg to about 25 mg each. Normally, such a dosage is given orally but parenteral administration may also be chosen.
  • the compounds of formula (I) above may be prepared by, or in analogy with, conventional methods. Formation of the central urethane linker is the key synthetic step in preparing the compounds of formula (I). A large number of activating reagents can be used for the formation of a urethane linker e.g. phosgene to form chloroformate of alcohols, or carbonyldiimidazole (CDI) to form imidazole carboxylates. Typically the urethane linkers incorporated into compounds of formula (I) have been synthesized utilizing bis-(4- nitrophenyl)carbonate as the activating agent, or by condensation of an alcohol with an isocyanate intermediate. The preparation of intermediates and compounds according to the examples of the present invention may in particular be illuminated by the following Schemes 1 and 2. Definitions of variables in the structures in the schemes herein are commensurate with those of corresponding positions in the formulae delineated herein.
  • R 1 , R 2 , R 3 and n are as defined in formula (I). - -
  • the formation of the urethane is typically a two step process but this may also be performed in a one-pot reaction by formation of the activated intermediate in situ.
  • a compound of formula (I) wherein R3 is H can be obtained in a single step by condensation of a (pyridine-4-yl)methanol derivative of formula (II) with the appropriate isocyanate of formula (V) in a suitable solvent (such as DCM), as shown in Scheme 2.
  • the necessary starting materials for preparing the compounds of formula (I) are either commercially available, or may be prepared by methods known in the art.
  • the processes described below in the experimental section may be carried out to give a compound of the invention in the form of a free base or as an acid addition salt.
  • a pharmaceutically acceptable acid addition salt may be obtained by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds. Examples of addition salt forming acids are mentioned above. - -
  • the compounds of formula (I) may possess one or more chiral carbon atoms, and they may therefore be obtained in the form of optical isomers, e.g., as a pure enantiomer, or as a mixture of enantiomers (racemate) or as a mixture containing diastereomers.
  • optical isomers e.g., as a pure enantiomer, or as a mixture of enantiomers (racemate) or as a mixture containing diastereomers.
  • the separation of mixtures of optical isomers to obtain pure enantiomers is well known in the art and may, for example, be achieved by fractional crystallization of salts with optically active (chiral) acids or by chromatographic separation on chiral columns.
  • the chemicals used in the synthetic routes delineated herein may include, for example, solvents, reagents, catalysts, and protecting group and deprotecting group reagents.
  • protecting groups are t-butoxycarbonyl (Boc), benzyl and trityl (triphenylmethyl).
  • the methods described above may also additionally include steps, either before or after the steps described specifically herein, to add or remove suitable protecting groups in order to ultimately allow synthesis of the compounds.
  • various synthetic steps may be performed in an alternate sequence or order to give the desired compounds.
  • Synthetic chemistry transformations and protecting group methodologies useful in synthesizing applicable compounds are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P.G.M.
  • Figure 1 is a schematic drawing illustrating weight gain and weight loss in mice during dark and light phases, respectively.
  • the graph illustrates the large nocturnal weight increase versus the comparatively small body weight change over 24 hours
  • Figure 2 shows the effect of Example 11 on the body weight in mice between the beginning of the dark phase and the beginning of the light phase (pm-am).
  • Figure 3 shows the effect of Example 15 on the body weight in mice between the beginning of the dark phase and the beginning of the light phase (pm-am).
  • Figure 4 shows the effect of Example 18 on the body weight in mice between the beginning of the dark phase and the beginning of the light phase (pm-am).
  • Figure 5 shows the concentration-dependent increase in [ 3 H] -thymidine incorporation by JEG-3 cells for leptin - -
  • System B Phenomenex Synergi Hydro RP, (150 x 4.6mm, 4 ⁇ m), gradient 0-20% CH 3 CN (+0.085% TFA) in H 2 O (+0.1% TFA), 1.5 mL/min, gradient time 7 min, 200-300 nm, 30 0 C; or System C: Phenomenex Synergi Hydro RP, (150 x 4.6mm, 4 ⁇ m), gradient 5-100% CH 3 CN in H 2 O (+0.1% HCO 2 H), 1.0 mL/min, gradient time 8 min, 30 °C;or
  • System E Phenomenex Synergi Hydro RP, (150 x 4.6mm, 4 ⁇ m), gradient 5-100% CH 3 CN (+0.085% TFA) in H 2 O (+0.1% TFA), 1.0 niL/min, gradient time 8 min, 30 °C; or System F: Phenomenex Synergi Hydro RP, (30 x 4.6mm, 4 ⁇ m), gradient 5-100% CH 3 CN (+0.085% TFA) in H 2 O (+0.1% TFA), 1.5 niL/min, gradient time 1.75 min, 30 °C.
  • mice gain about 1 g in weight during the dark phase and then loose the majority of this weight gain during the light phase, as represented in Figure 1.
  • the weight difference over any 24 hour period is very small whilst the weight difference between the beginning of the dark phase and the beginning of the light phase (pm-am) is maximal.
  • mice are dosed with an active compound on two consecutive days and the bodyweight change is recorded 48 hours after the first dose then no significant effect is observed. However if the body weight change over the dark phase only is considered a significant and robust effect is seen. This is because the mice rebound during the light phase to compensate for the lack of weight gain over the dark phase. Very active long lasting compounds may also diminish this rebound and reduce the body weight over the 48 hours.
  • the weight difference between the beginning of the dark phase and the beginning of the light phase (pm-am) is greater than the weight difference measured between pm and pm on 2 consecutive days.
  • the effect of the compounds on the pm-am difference was therefore studied in order to maximise the effect window.
  • mice C57 bl/6 mice were grouped (5 per cage) and left 5 days for acclimatisation.
  • a single intraperitoneally (ip) administered dose 60 mg/kg was given just prior to the dark phase.
  • Compounds were either water soluble or dissolved in up to 3% cremophor (in this case the vehicle also contained cremophor).
  • the pH was adjusted from a minimum of 5.5 to a maximum of 8 depending on the nature of the compound.
  • leptin receptor expression in non-recombinant system is often fluctuating and care must be given to identify a system where signal stability remains within experiments.
  • leptin receptor antagonist mimetics could be identified by evaluating their action vs. leptin (see below).
  • Leptin is produced chiefly in adipose cells, but in humans, mRNA encoding leptin is also present in the placenta. Here, leptin might play an important proliferative role in the microvasculature. The possibility to use this hypothesis in a native cell line was evaluated.
  • JEG-3 cells choriocarcinoma cell line
  • leptin is able to stimulate proliferation up to 3 fold (Biol. Reprod. (2007) 76: 203-10).
  • the radioactivity incorporated by the cells is an index of their proliferative activity and is measured in counts per minute (CPM) with a liquid scintillation beta counter.
  • This approach has the advantage of using a non-recombinant system and has reasonable reproducibility and robustness.
  • test species is given a bolus dose of the substrate under investigation, usually via intravenous (IV) or oral (PO) routes.
  • IV intravenous
  • PO oral
  • blood samples are taken and the resultant plasma extracted and analysed for substrate concentration and, where appropriate, metabolite concentration.
  • animals from another group are sacrificed, brains isolated and the brain surface cleaned. Brain samples are then homogenised, extracted and analysed for substrate concentration and, where appropriate, metabolite concentration.
  • microdialysis probes are implanted into one or more brain regions of the test species and samples collected at appropriate time points for subsequent analysis. This method has the advantage of measuring only extra-cellular substrate concentration. Plasma and brain concentrations are then compared and ratios calculated, either by comparison of averaged concentrations at individual time points, or by calculation of the area-under-the-curve (AUC) of the concentration-time plots.
  • AUC area-under-the-curve

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Abstract

The present invention relates to new compounds of formula (I), to pharmaceutical compositions comprising these compounds and to the use of these compounds as leptin receptor modulator mimetics in the preparation of medicaments against conditions associated with weight gain, type 2 diabetes and dyslipidemias.

Description

NEW PYRIDINE DERIVATIVES AS LEPTIN RECEPTOR MODULATOR MIMETICS
FIELD OF THE INVENTION
The present application relates to new pyridine derivatives, to pharmaceutical compositions comprising these compounds and to the use of these compounds as leptin receptor modulator mimetics in the preparation of medicaments against conditions associated with weight gain, type 2 diabetes and dyslipidemias.
BACKGROUND ART
The prevalence of obesity is increasing in the industrialized world. Typically, the first line of treatment is to offer diet and life style advice to patients, such as reducing the fat content of their diet and increasing their physical activity. However, some patients may also need to undergo drug therapy to maintain the beneficial results obtained from adapting the aforementioned diet and lifestyle changes.
Leptin is a hormone synthesized in fat cells that is believed to act in the hypothalamus to reduce food intake and body weight (see, e.g., Bryson, J. M. (2000) Diabetes, Obesity and Metabolism 2: 83-89).
It has been shown that in obese humans the ratio of leptin in the cerebrospinal fluid to that of circulating leptin is decreased (Koistinen et al, (1998) Eur. J. Clin. Invest. 28: 894-897). This suggests that the capacity for leptin transport into the brain is deficient in the obese state. Indeed, in animal models of obesity (NZO mouse and Koletsky rat), defects in leptin transport have been shown to result in reduced brain leptin content (Kastin, A. J. (1999) Peptides 20: 1449-1453; Banks, W. A. et al, (2002) Brain Res. 950: 130-136). In studies involving dietary- induced obese rodents (a rodent model that is believed to more closely resemble human obesity, see, e.g., Van Heek et al. (1997) J. Clin. Invest. 99: 385-390), excess leptin administered peripherally was shown to be ineffective in reducing food intake and body weight, whereas leptin injected directly into the brain was effective in reducing food intake and body weight. It has also been shown that in obese humans with excess circulating leptin, the signaling system became desensitized to the continual stimulation of the leptin receptors (Mantzoros, C. S. (1999) Ann. Intern. Med. 130: 671-680). - -
Amgen has conducted clinical trials with recombinant methionyl human leptin. The results from these trials were mixed, as even in the presence of high plasma concentrations of leptin weight loss was variable, and the average weight reduction in the cohort of patients tested relatively small (Obesity Strategic Perspective, Datamonitor, 2001). Several attempts at finding active fragments have been reported in the literature since the discovery of the leptin gene coding sequence. An example is by Samson et al. (1996) Endocrinol. 137: 5182-5185 which describes an active fragment at the N-terminal (22 to 56). This sequence was shown to reduce food intake when injected ICV whereas a sequence taken at the C-terminal was shown not to have any effect. Leptin fragments are also disclosed in International Patent Application WO 97/46585.
Other reports looking at the C-terminus part of the sequence reported a possible stimulation of luteinising hormone production by a 116-130 fragment (Gonzalez et al., (1999) Neuroendocrinology 70:213-220) and an effect on GH production following GHRH administration (fragment 126-140) (Hanew (2003) Eur. J. Endocrin. 149: 407-412). Leptin has recently been associated with inflammation. It has been reported that circulating leptin levels rise during bacterial infection and in inflammation (see Otero, M et al. (2005) FEBS Lett. 579: 295-301 and references therein). Leptin can also act to increase inflammation by enhancing the release of pro -inflammatory cytokines TNF and IL-6 from inflammatory cells (Zarkesh-Esfahani, H. et al. (2001) J. Immunol. 167: 4593-4599). These agents in turn can contribute to the insulin resistance commonly seen in obese patients by reducing the efficacy of insulin receptor signaling (Lyon, C. J. et al. (2003) Endocrinol. 44: 2195-2200). Continuous low grade inflammation is believed to be associated with obesity (in the presence and absence of insulin resistance and Type II diabetes) (Browning et al. (2004) Metabolism 53: 899-903, Inflammatory markers elevated in blood of obese women; Mangge et al. (2004) Exp. Clin. Endocrinol. Diabetes 112: 378- 382, Juvenile obesity correlates with serum inflammatory marker C-reactive protein; Maachi et al. (2004) Int. J. Obes. Relat. Metab. Disord. 28: 993-997, Systemic low grade inflammation in obese people). Leptin has also been implicated in the process of atherogenesis, by promoting lipid uptake into macrophages and endothelial dysfunction, thus promoting the formation of atherosclerotic plaques (see Lyon, C. J. et al. (2003) Endocrinol. 144: 2195-2200).
Leptin has also been shown to promote the formation of new blood vessels (angiogenesis) a process implicated in the growth of adipose tissue (Bouloumie A, et al. (1998) Circ. Res. - -
83: 1059-1066). Angiogenesis has also been implicated in diabetic retinopathy (Suganami, E. et al. (2004) Diabetes. 53: 2443-2448).
Angiogenesis is also believed to be involved with the growth of new blood vessels that feed abnormal tumour cells. Elevated leptin levels have been associated with a number of cancers, in particular breast, prostate and gastrointestinal cancers in humans (Somasundar P. et al. (2004) J. Surg. Res. 116: 337-349).
Leptin receptor agonists may also be used in the manufacture of a medicament to promote wound healing (Gorden, P. and Gavrilova, O. (2003) Current Opinion in Pharmacology 3: 655-659). Further, it has been shown that elevating leptin signaling in the brain may represent an approach for the treatment of depressive disorders (Lu, Xin-Yun et al. (2006) PNAS 103: 1593-1598).
DISCLOSURE OF THE INVENTION
It has surprisingly been found that compounds of formula (I) are effective in reducing body weight and food intake in rodents. While not wishing to be bound by theory, it is proposed that the compounds of formula I modulate the leptin receptor signaling pathway.
In some embodiments, compounds with leptin receptor agonistic like properties can be useful for the treatment of disorders relating to leptin signaling, as well as conditions associated with weight gain, such as obesity. The inventors hypothesized that small molecule CNS penetrant leptin mimetics would be able to by-pass the limiting uptake system into the brain. Further, assuming that this situation mirrors the human obese condition, the inventors believe that a CNS-penetrant leptinoid with a relatively long duration of action would make an effective therapy for the obese state and its attendant complications, in particular (but not limited to) diabetes.
In other embodiments, compounds with leptin receptor antagonistic like properties could be useful for the treatment of inflammation, atherosclerosis, diabetic retinopathy and nephropathy. - -
In a first aspect, the disclosure relates to a compound of formula (I),
or a pharmaceutically acceptable salt, solvate, hydrate, geometrical isomer, tautomer or optical isomer thereof, wherein:
each R1 is independently selected from Ci_4-alkyl, Ci_4-alkoxy, halogen, cyano and CF3;
R2 is Ci_6-alkyl (optionally substituted with one or more substituents selected from hydroxy, halogen and cyano) or -[C(R4A)(R4B)]m-R5;
R3 is hydrogen, Ci_4-alkyl or fluoro-Ci_4-alkyl;
R4A and R4B are each independently selected from hydrogen, halogen, hydroxy, Ci_4-alkyl, fluoro-Ci_4-alkyl and hydroxy-Ci_4-alkyl;
R5 is C3-8-cycloalkyl, Cβ-io-aryl, heterocyclyl or heteroaryl, each of which is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro,
CF3 and Ci_4-alkyl;
m is 0, 1 or 2; and n is 0, 1, 2, 3 or 4;
with the proviso that the compound is not selected from:
• pyridin-4-ylmethyl bis(2-chloroethyl)carbamate;
• (2,6-dichloropyridin-4-yl)methyl dimethylcarbamate;
• (2,6-dichloropyridin-4-ylmethyl propylcarbamate; • pyridin-4-ylmethyl methylcarbamate;
• pyridin-4-ylmethyl isopropylcarbamate;
• pyridin-4-ylmethyl [3 -(trifluoromethyl)phenyl] carbamate;
• pyridin-4-ylmethyl (5-chloro-2-methoxyphenyl)carbamate;
• pyridin-4-ylmethyl (2-methoxyphenyl)carbamate; • pyridin-4-ylmethyl (2,6-dimethylphenyl)carbamate; - -
• pyridin-4-ylmethyl (2,4,6-trimethylphenyl)carbamate;
• pyridin-4-ylmethyl (5-chloro-2,4-dimethoxyphenyl)carbamate;
• pyridin-4-ylmethyl (2-methyl-5-nitrophenyl)carbamate;
• pyridin-4-ylmethyl (3-ethylphenyl)carbamate; • pyridin-4-ylmethyl (2,4-dimethylphenyl)carbamate;
• pyridin-4-ylmethyl (3,4,5-trimethoxyphenyl)carbamate;
• pyridin-4-ylmethyl cyclohexyl(methyl)carbamate;
• pyridin-4-ylmethyl pyridin-3-ylcarbamate;
• pyridin-4-ylmethyl (4-methoxyphenyl)carbamate; • pyridin-4-ylmethyl (2,6-dichlorophenyl)carbamate;
• pyridin-4-ylmethyl cyclohexylcarbamate;
• pyridin-4-ylmethyl pyridin-4-ylcarbamate;
• pyridin-4-ylmethyl (4-fluorophenyl)carbamate;
• pyridin-4-ylmethyl [(2-chlorophenyl)methyl] carbamate; • pyridin-4-ylmethyl (3-nitrophenyl)carbamate;
• pyridin-4-ylmethyl (3,5-dimethoxyphenyl)carbamate;
• pyridin-4-ylmethyl (4-methylphenyl)carbamate;
• pyridin-4-ylmethyl (3-chlorophenyl)carbamate;
• pyridin-4-ylmethyl (4-chlorophenyl)carbamate; • (2,6-dichloropyridin-4-yl)methyl phenylcarbamate;
• (2,6-dimethylpyridin-4-yl)methyl phenylcarbamate;
• pyridin-4-ylmethyl phenylcarbamate; and
• pyridin-4-ylmethyl (3,4-dimethoxyphenyl)carbamate.
In a preferred embodiment, R2 is Ci_6-alkyl which is substituted with one or more substituents selected from hydroxy, halogen and cyano, more preferably hydroxy. In a yet more preferred embodiment, R2 is 2-hydroxyethyl, l-(hydroxymethyl)-2-methylpropyl or 1 -(hydroxymethyl)-3 -methylbutyl.
In another preferred embodiment, R2 is -[C(R4A)(R4B)]m-R5, wherein R5 is phenyl, C3-6- cycloalkyl, tetrahydrofuranyl, indanyl or isoxazolyl, and is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro, CF3 or Ci_4-alkyl. In a - -
more preferred embodiment, R5 is cyclopropyl, cyclopentyl, 1-hydroxycyclohexyl, 2-tetra- hydrofuranyl, 3-tetrahydrofuranyl, 2-hydroxy-lH-inden-l-yl or 3,5-dimethylisoxazol-4-yl.
m is preferably 0 or 1. When m is 1, R4A and R4B are preferably independently selected from hydrogen, methyl and hydroxymethyl.
R3 is preferably hydrogen or methyl.
Specific preferred compounds according to the disclosure are those selected from the group consisting of:
• pyridin-4-ylmethyl dimethylcarbamate;
• pyridin-4-ylmethyl [(2S)-tetrahydrofuran-2-ylmethyl]carbamate;
• pyridin-4-ylmethyl (2-hydroxyethyl)carbamate; • pyridin-4-ylmethyl (2-hydroxyethyl)methylcarbamate;
• pyridin-4-ylmethyl [(IR)- 1 -(hydroxymethyl)-2-methylpropyl]carbamate;
• pyridin-4-ylmethyl [(IR)- 1 -(hydroxymethyl)-3-methylbutyl]carbamate;
• pyridin-4-ylmethyl cyclopentylcarbamate;
• pyridin-4-ylmethyl (3i?)-tetrahydrofuran-3-ylcarbamate; • pyridin-4-ylmethyl [(l-hydroxycyclohexy^methyljcarbamate;
• (pyridin-4-yl)methyl (li?,25)-2,3-dihydro-2-hydroxy- lH-inden- 1 -ylcarbamate;
• pyridin-4-ylmethyl [(15)-l-phenylethyl]carbamate;
• pyridin-4-ylmethyl [(li?)-2-hydroxy-l-phenylethyl]carbamate;
• (2,6-dimethylpyridin-4-yl)methyl (1 -methyl- 1 -phenylethyl)carbamate; • (2,6-dimethylpyridin-4-yl)methyl tert-butylcarbamate;
• (2,6-dimethylpyridin-4-yl)methyl cyclopentylcarbamate;
• (2,6-dimethylpyridin-4-yl)methyl (cyclopropylmethyl)carbamate;
• (2,6-dimethylpyridin-4-yl)methyl (3i?)-tetrahydrofuran-3-ylcarbamate; and
• (2,6-dimethylpyridin-4-yl)methyl (3,5-dimethylisoxazol-4-yl)carbamate.
Another aspect of the present disclosure is a compound of formula (I) for use in therapy. - -
In a further aspect, the invention relates to a compound of formula (I) for use in the treatment or prevention of any of the disorders or conditions described herein.
In a yet further aspect, the invention relates to the use of the compounds of formula (I) in the manufacture of a medicament for the treatment or prevention of any of the disorders or conditions described herein.
In some embodiments, said compounds may be used for the treatment or prevention of a condition that is prevented, treated, or ameliorated by selective action via the leptin receptor.
In some embodiments, compounds of formula (I) may be used for the treatment or prevention of conditions (in particular, metabolic conditions) that are associated with weight gain. Conditions associated with weight gain include diseases, disorders, or other conditions that have an increased incidence in obese or overweight subjects. Examples include: lipodystrophy, HIV lipodystrophy, diabetes (type 2), insulin resistance, metabolic syndrome, hyperglycemia, hyperinsulinemia, dyslipidemia, hepatic steatosis, hyperphagia, hypertension, hypertriglyceridemia, infertility, a skin disorder associated with weight gain, macular degeneration. In some embodiments, compounds of the invention may also be used in the manufacture of a medicament for maintaining weight loss of a subject.
In some embodiments, compounds of formula (I) which are leptin receptor agonist mimetics may also be used to promote wound healing.
In some embodiments, compounds of formula (I) which are leptin receptor agonist mimetics may also be used for the treatment or prevention of conditions that cause a decrease in circulating leptin concentrations, and the consequent malfunction of the immune and reproductive systems. Examples of such conditions and malfunctions include severe weight loss, dysmenorrhea, amenorrhea, female infertility, immunodeficiency and conditions associated with low testosterone levels. - -
In some embodiments, compounds of formula (I) which are leptin receptor agonist mimetics may also be used for the treatment or prevention of conditions caused as a result of leptin deficiency, or a leptin or leptin receptor mutation.
In some other embodiments, compounds of formula (I) which are leptin receptor antagonist mimetics may be used for the treatment or prevention of inflammatory conditions or diseases, low level inflammation associated with obesity and excess plasma leptin and in reducing other complications associated with obesity including atherosclerosis, and for the correction of insulin resistance seen in Metabolic Syndrome and diabetes.
In some embodiments, compounds of formula (I) which are leptin receptor antagonist mimetics can be used for the treatment or prevention of inflammation caused by or associated with: cancer (such as leukemias, lymphomas, carcinomas, colon cancer, breast cancer, lung cancer, pancreatic cancer, hepatocellular carcinoma, kidney cancer, melanoma, hepatic, lung, breast, and prostate metastases, etc.); auto-immune disease (such as organ transplant rejection, lupus erythematosus, graft v. host rejection, allograft rejections, multiple sclerosis, rheumatoid arthritis, type I diabetes mellitus including the destruction of pancreatic islets leading to diabetes and the inflammatory consequences of diabetes); autoimmune damage (including multiple sclerosis, Guillam Barre Syndrome, myasthenia gravis); cardiovascular conditions associated with poor tissue perfusion and inflammation (such as atheromas, atherosclerosis, stroke, ischaemia-reperfusion injury, claudication, spinal cord injury, congestive heart failure, vasculitis, haemorrhagic shock, vasospasm following subarachnoid haemorrhage, vasospasm following cerebrovascular accident, pleuritis, pericarditis, the cardiovascular complications of diabetes); ischaemia- reperfusion injury, ischaemia and associated inflammation, restenosis following angioplasty and inflammatory aneurysms; epilepsy, neurodegeneration (including Alzheimer's Disease), arthritis (such as rheumatoid arthritis, osteoarthritis, rheumatoid spondylitis, gouty arthritis), fibrosis (for example of the lung, skin and liver), multiple sclerosis, sepsis, septic shock, encephalitis, infectious arthritis, Jarisch-Herxheimer reaction, shingles, toxic shock, cerebral malaria, Lyme's disease, endotoxic shock, gram negative shock, haemorrhagic shock, hepatitis (arising both from tissue damage or viral infection), deep vein thrombosis, gout; conditions associated with breathing difficulties (e.g. chronic obstructive pulmonary disease, impeded and obstructed airways, - -
bronchoconstriction, pulmonary vasoconstriction, impeded respiration, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcosis, cystic fibrosis, pulmonary hypertension, pulmonary vasoconstriction, emphysema, bronchial allergy and/or inflammation, asthma, hay fever, rhinitis, vernal conjunctivitis and adult respiratory distress syndrome); conditions associated with inflammation of the skin (including psoriasis, eczema, ulcers, contact dermatitis); conditions associated with inflammation of the bowel (including Crohn's disease, ulcerative colitis and pyresis, irritable bowel syndrome, inflammatory bowel disease); HIV (particularly HIV infection), cerebral malaria, bacterial meningitis, osteoporosis and other bone resorption diseases, osteoarthritis, infertility from endometriosis, fever and myalgia due to infection, and other conditions mediated by excessive anti-inflammatory cell (including neutrophil, eosinophil, macrophage and T- cell) activity.
In some embodiments, compounds of formula (I) which are leptin receptor antagonists mimetics may be used for the treatment or prevention of macro or micro vascular complications of type 1 or 2 diabetes, retinopathy, nephropathy, autonomic neuropathy, or blood vessel damage caused by ischaemia or atherosclerosis.
In some embodiments, compounds of formula (I) which are leptin receptor antagonist mimetics may be used to inhibit angiogenesis. Compounds that inhibit angiogenesis may be used for the treatment or prevention of obesity or complications associated with obesity. Compounds that inhibit angiogenesis may be used for the treatment or prevention of complications associated with inflammation diabetic retinopathy, or tumour growth particularly in breast, prostate or gastrointestinal cancer.
In a further aspect, the disclosure relates to a method for the treatment or prevention of any of the disorders or conditions described herein, which includes administering to a subject (e.g., a subject in need thereof, e.g., a mammal) an effective amount of a compound of formula I.
Methods delineated herein include those wherein the subject is identified as in need of a particular stated treatment. Identifying a subject in need of such treatment can be in the - -
judgment of a subject or a health care professional and can be subjective (e.g. opinion) or objective (e.g. measurable by a test or diagnostic method).
In other aspects, the methods herein include those further comprising monitoring subject response to the treatment administrations. Such monitoring may include periodic sampling of subject tissue, fluids, specimens, cells, proteins, chemical markers, genetic materials, etc. as markers or indicators of the treatment regimen. In other methods, the subject is prescreened or identified as in need of such treatment by assessment for a relevant marker or indicator of suitability for such treatment. In one embodiment, the invention provides a method of monitoring treatment progress. The method includes the step of determining a level of diagnostic marker (Marker) (e.g., any target or cell type delineated herein modulated by a compound herein) or diagnostic measurement (e.g., screen, assay) in a subject suffering from or susceptible to a disorder or symptoms thereof delineated herein, in which the subject has been administered a therapeutic amount of a compound herein sufficient to treat the disease or symptoms thereof. The level of Marker determined in the method can be compared to known levels of Marker in either healthy normal controls or in other afflicted patients to establish the subject's disease status. In preferred embodiments, a second level of Marker in the subject is determined at a time point later than the determination of the first level, and the two levels are compared to monitor the course of disease or the efficacy of the therapy. In certain preferred embodiments, a pre-treatment level of Marker in the subject is determined prior to beginning treatment according to this invention; this pre-treatment level of Marker can then be compared to the level of Marker in the subject after the treatment commences, to determine the efficacy of the treatment. In certain method embodiments, a level of Marker or Marker activity in a subject is determined at least once. Comparison of Marker levels, e.g., to another measurement of Marker level obtained previously or subsequently from the same patient, another patient, or a normal subject, may be useful in determining whether therapy according to the disclosure is having the desired effect, and thereby permitting adjustment of dosage levels as appropriate. Determination of Marker levels may be performed using any suitable sampling/expression assay method known in the art or described herein. Preferably, a tissue or fluid sample is first removed from a subject. Examples of suitable samples include blood, urine, tissue, mouth or cheek cells, and hair samples containing roots. Other suitable samples would be known to the person skilled in the art. Determination of protein - -
levels and/or mRNA levels (e.g., Marker levels) in the sample can be performed using any suitable technique known in the art, including, but not limited to, enzyme immunoassay, ELISA, radio labeling/assay techniques, blotting/chemiluminescence methods, real-time PCR, and the like.
In some embodiments, it may be advantageous if a compound of formula (I) is able to penetrate the central nervous system. In other embodiments, it may be advantageous if a compound of formula (I) is not able to penetrate the CNS. In general, it is expected that compounds that are leptin receptor agonist mimetics may be particularly useful for the treatment or prevention of obesity, insulin resistance, or diabetes (particularly glucose intolerance) if these compounds can penetrate the CNS. A person of ordinary skill in the art can readily determine whether a compound can penetrate the CNS. A suitable method that may be used is described in the Biological Methods section.
A leptin receptor response may be measured in any suitable way. In vitro, this may be done be measuring leptin receptor signaling. For example, phosphorylation of Akt, STAT3, STAT5, MAPK, shp2 or the leptin receptor in response to binding of leptin or a compound of the invention to the leptin receptor may be measured. The extent of phosphorylation of Akt, STAT3, STAT5, MAPK, shp2 or the leptin receptor may be determined for example by Western blotting or by ELISA. Alternatively, a STAT reporter assay may be used, for example STAT driven luciferase expression. A cell line expressing the leptin receptor may be used for such assays. In vivo, leptin receptor response may be measured by determining the reduction in food intake and body weight after administration of leptin or a compound of formula (I).
The Biological Methods below describe assays and methods that can be used to determine whether a compound of formula (I) is a leptin receptor agonist mimetic or a leptin receptor antagonist mimetic.
A compound of formula (I) may be administered with or without other therapeutic agents. For example, where it is desired to reduce inflammation, a compound may be administered with an anti-inflammatory agent (for example, disease modifying anti-rheumatic drugs such as methotrexate, sulphasalazine and cytokine inactivating agents, steroids, NSAIDs, - -
cannabinoids, tachykinin modulators, or bradykinin modulators). Where it is desired to provide an anti-tumour effect, a compound may be administered with a cytotoxic agent (for example, methotrexate, cyclophosphamide) or another anti-tumour drug.
Compounds of formula (I) may be radiolabeled (for example with tritium or radioactive iodine) for in vitro or in vivo applications, such as receptor displacement studies or receptor imaging.
DEFINITIONS
The following definitions shall apply throughout the specification and the appended claims.
Unless otherwise stated or indicated, the term "Ci_6-alkyl" denotes a straight or branched alkyl group having from 1 to 4 carbon atoms. Examples of said Ci_6-alkyl include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl and t-butyl, as well as straight- and branched-chain pentyl and hexyl. For parts of the range "Ci_6-alkyl" all subgroups thereof are contemplated such as Ci_5-alkyl, Ci_4-alkyl, Ci_3-alkyl, Ci_2-alkyl, C2-6-alkyl, C2-5-alkyl, C2_4-alkyl, C2_3-alkyl, C3-6-alkyl, C3-5-alkyl,C3-4-alkyl, C4-6-alkyl, C4-5-alkyl and C5.6-alkyl. Unless otherwise stated or indicated, the term "hydroxy-Ci_4-alkyl" denotes a straight or branched Ci_4-alkyl group that has a hydrogen atom thereof replaced with OH. Examples of said hydroxy-Ci_4-alkyl include hydroxymethyl, 2-hydroxyethyl and 2-hydroxypropyl. Unless otherwise stated or indicated, the term "fluoro-Ci_4-alkyl" denotes a straight or branched Ci_4-alkyl group substituted by one or more fluorine atoms. Examples of said fluoro-Ci_4-alkyl include fluoromethyl, trifluoromethyl, 2-fluoroethyl and 2,2,2- trifluoroethyl.
Unless otherwise stated or indicated, the term "Ci_4-alkoxy" denotes a straight or branched alkoxy group having from 1 to 4 carbon atoms. Examples of said Ci_4-alkoxy include methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, iso-butoxy, sec-butoxy and t-butoxy. For parts of the range "Ci_4-alkoxy" all subgroups thereof are contemplated such as C1-3- alkoxy, Ci_2-alkoxy, C2-4-alkoxy, C2-3-alkoxy and C3_4-alkoxy.
Unless otherwise stated or indicated, the term "C3_8-cycloalkyl" denotes a mono- or bicyclic saturated hydrocarbon ring system having 3 to 8 carbon atoms. Bicyclic ring systems can be either fused or bridged. In a bridged cycloalkyl ring system, two non- - -
adjacent carbon atoms of a monocyclic ring are linked by an alkylene bridge of between one and three additional carbon atoms. Examples of C3_8-cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, as well as bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl and bicyclo[3.2.1]octyl. For parts of the range "Cs-s-cycloalkyl" all subgroups thereof are contemplated such as C3-7-cycloalkyl, C3_6-cycloalkyl, C3_5-cycloalkyl, C3_4-cycloalkyl, C4_8-cycloalkyl, C4_7-cycloalkyl, C4_6-cycloalkyl, C4_5-cycloalkyl, Cs_8-cycloalkyl, Cs_7-cycloalkyl, Cs_6-cycloalkyl, C6-8-cycloalkyl and C6-7-cycloalkyl. Unless otherwise stated or indicated, the term "Cβ-io-aryl" refers to a mono- or bicyclic hydrocarbon ring system comprising 6 to 10 ring atoms and wherein at least one ring is an aromatic ring. Examples of Cβ-io-aryl include phenyl, indenyl, 2,3-dihydroindenyl (indanyl), 1-naphthyl, 2-naphthyl or 1,2,3,4-tetrahydronaphthyl.
Unless otherwise stated or indicated, the term "heterocyclyl" refers to a stable, fully saturated or partially unsaturated mono- or bicyclic ring system having 3 to 8 ring atoms with at least one heteroatom such as O, N, or S, and the remaining ring atoms are carbon. Examples of heterocyclyl include piperidinyl, tetrahydropyranyl, tetrahydrofuranyl, azepinyl, azetidinyl, pyrrolidinyl, morpholinyl, imidazolinyl, thiomorpholinyl, pyranyl, dioxanyl, piperazinyl, as well as l-azabicyclo[2.2.2]octane, l-azabicyclo[2.2.1]heptane and azabicyclo[2.2.2]oct-2-ene. When present, a sulfur atom may be in an oxidized form (i.e., S=O or O=S=O). An exemplary heterocyclic group containing sulfur in oxidized form is thiomorpholine- 1 , 1 -dioxide.
Unless otherwise stated or indicated, the term "heteroaryl" refers to a mono- or bicyclic heteroaromatic ring system having 5 to 10 ring atoms in which one or more of the ring atoms are other than carbon, such as nitrogen, sulphur or oxygen. Only one ring need to be aromatic and said heteroaryl moiety can be linked to the remainder of the molecule via a carbon or nitrogen atom in any ring. Examples of heteroaryl include furyl, pyrrolyl, thienyl, oxazolyl, isoxazolyl, imidazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, tetrazolyl, quinazolinyl, indolyl, isoindolyl, 1,3-dihydro-isoindolyl, pyrazolyl, pyridazinyl, quinolinyl, quinoxalinyl, thiadiazolyl, benzofuranyl, 2,3-dihydrobenzofuranyl, 1,3- benzodioxolyl, 1 ,4-benzodioxinyl, 2,3-dihydro-l,4-benzodioxinyl, benzothiazolyl, benzimidazolyl, benzothiadiazolyl, benzotriazolyl, indolinyl, isoindolinyl and chromanyl. "Halogen" refers to fluorine, chlorine, bromine or iodine. "Hydroxy" refers to the -OH radical. - -
"Nitro" refers to the -NO2 radical. "Cyano" refers to the -CN radical.
"Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not.
The term "mammal" includes organisms, which include mice, rats, cows, sheep, pigs, rabbits, goats, and horses, monkeys, dogs, cats, and preferably humans. The subject may be a human subject or a non human animal, particularly a domesticated animal, such as a dog. "Pharmaceutically acceptable" means being useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable and includes being useful for veterinary use as well as human pharmaceutical use.
"Treatment" as used herein includes prophylaxis of the named disorder or condition, or amelioration or elimination of the disorder once it has been established. "An effective amount" refers to an amount of a compound that confers a therapeutic effect (e.g., treats, controls, ameliorates, prevents, delays the onset of, or reduces the risk of developing a disease, disorder, or condition or symptoms thereof) on the treated subject. The therapeutic effect may be objective (i.e., measurable by some test or marker) or subjective (i.e., subject gives an indication of or feels an effect). "Prodrugs" refers to compounds that may be converted under physiological conditions or by so lvo lysis to a biologically active compound of formula (I). A prodrug may be inactive when administered to a subject in need thereof, but is converted in vivo to an active compound of formula (I). Prodrugs are typically rapidly transformed in vivo to yield the parent compound, e.g. by hydrolysis in the blood. The prodrug compound usually offers advantages of solubility, tissue compatibility or delayed release in a mammalian organism (see Silverman, R. B., The Organic Chemistry of Drug Design and Drug Action, 2nd Ed., Elsevier Academic Press (2004), pp. 498-549). Prodrugs may be prepared by modifying functional groups, such as a hydroxy, amino or mercapto groups, present in a compound of formula (I) in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compound. Examples of prodrugs include, but are not limited to, acetate, formate and succinate derivatives of hydroxy functional groups or phenyl carbamate derivatives of amino functional groups. - -
Throughout the specification and the appended claims, a given chemical formula or name shall also encompass all salts, hydrates, solvates, N-oxides and prodrug forms thereof. Further, a given chemical formula or name shall encompass all tautomeric and stereoisomeric forms thereof. Stereoisomers include enantiomers and diastereomers. Enantiomers can be present in their pure forms, or as racemic (equal) or unequal mixtures of two enantiomers. Diastereomers can be present in their pure forms, or as mixtures of diastereomers. Diastereomers also include geometrical isomers, which can be present in their pure cis or trans forms or as mixtures of those. The compounds of formula (I) may be used as such or, where appropriate, as pharmacologically acceptable salts (acid or base addition salts) thereof. The pharmacologically acceptable addition salts mentioned below are meant to comprise the therapeutically active non-toxic acid and base addition salt forms that the compounds are able to form. Compounds that have basic properties can be converted to their pharmaceutically acceptable acid addition salts by treating the base form with an appropriate acid. Exemplary acids include inorganic acids, such as hydrogen chloride, hydrogen bromide, hydrogen iodide, sulphuric acid, phosphoric acid; and organic acids such as formic acid, acetic acid, propanoic acid, hydroxyacetic acid, lactic acid, pyruvic acid, glycolic acid, maleic acid, malonic acid, oxalic acid, benzenesulphonic acid, toluenesulphonic acid, methanesulphonic acid, trifluoroacetic acid, fumaric acid, succinic acid, malic acid, tartaric acid, citric acid, salicylic acid, /^-aminosalicylic acid, pamoic acid, benzoic acid, ascorbic acid and the like. Exemplary base addition salt forms are the sodium, potassium, calcium salts, and salts with pharmaceutically acceptable amines such as, for example, ammonia, alkylamines, benzathine, and amino acids, such as, e.g. arginine and lysine. The term addition salt as used herein also comprises solvates which the compounds and salts thereof are able to form, such as, for example, hydrates, alcoholates and the like.
COMPOSITIONS
For clinical use, the compounds of formula (I) are formulated into pharmaceutical formulations for various modes of administration. It will be appreciated that the compounds may be administered together with a physiologically acceptable carrier, excipient, or diluent. The pharmaceutical compositions may be administered by any - -
suitable route, preferably by oral, rectal, nasal, topical (including buccal and sublingual), sublingual, transdermal, intrathecal, transmucosal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration.
Other formulations may conveniently be presented in unit dosage form, e.g., tablets and sustained release capsules, and in liposomes, and may be prepared by any methods well known in the art of pharmacy. Pharmaceutical formulations are usually prepared by mixing the active substance, or a pharmaceutically acceptable salt thereof, with conventional pharmaceutically acceptable carriers, diluents or excipients. Examples of excipients are water, gelatin, gum arabicum, lactose, microcrystalline cellulose, starch, sodium starch glycolate, calcium hydrogen phosphate, magnesium stearate, talcum, colloidal silicon dioxide, and the like. Such formulations may also contain other pharmacologically active agents, and conventional additives, such as stabilizers, wetting agents, emulsifiers, flavouring agents, buffers, and the like. Usually, the amount of active compounds is between 0.1-95% by weight of the preparation, preferably between 0.2-20% by weight in preparations for parenteral use and more preferably between 1-50% by weight in preparations for oral administration.
The formulations can be further prepared by known methods such as granulation, compression, microencapsulation, spray coating, etc. The formulations may be prepared by conventional methods in the dosage form of tablets, capsules, granules, powders, syrups, suspensions, suppositories or injections. Liquid formulations may be prepared by dissolving or suspending the active substance in water or other suitable vehicles. Tablets and granules may be coated in a conventional manner. To maintain therapeutically effective plasma concentrations for extended periods of time, the compounds may be incorporated into slow release formulations. The dose level and frequency of dosage of the specific compound will vary depending on a variety of factors including the potency of the specific compound employed, the metabolic stability and length of action of that compound, the patient's age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the condition to be treated, and the patient undergoing therapy. The daily dosage may, for example, range from about 0.001 mg to about 100 mg per kilo of body weight, administered singly or multiply in doses, e.g. from about 0.01 mg to about 25 mg each. Normally, such a dosage is given orally but parenteral administration may also be chosen. - -
PREPARATION OF COMPOUNDS OF THE INVENTION
The compounds of formula (I) above may be prepared by, or in analogy with, conventional methods. Formation of the central urethane linker is the key synthetic step in preparing the compounds of formula (I). A large number of activating reagents can be used for the formation of a urethane linker e.g. phosgene to form chloroformate of alcohols, or carbonyldiimidazole (CDI) to form imidazole carboxylates. Typically the urethane linkers incorporated into compounds of formula (I) have been synthesized utilizing bis-(4- nitrophenyl)carbonate as the activating agent, or by condensation of an alcohol with an isocyanate intermediate. The preparation of intermediates and compounds according to the examples of the present invention may in particular be illuminated by the following Schemes 1 and 2. Definitions of variables in the structures in the schemes herein are commensurate with those of corresponding positions in the formulae delineated herein.
Scheme 1
,Ff
HN'
(IV)
wherein R1, R2, R3 and n are as defined in formula (I). - -
Compounds of formula (I) can easily be prepared in only a few synthetic steps. Activation of a (pyridine-4-yl)methanol derivative of formula (II) with δώ-(4-nitrophenyl)carbonate in the presence of a base (such as NMM) in an aprotic solvent (such as DCM) results in the corresponding carbonate of formula (III). Treatment of (III) with the appropriate amine (IV) in the presence of a base (such as DIPEA) and an activating agent (such as DMAP) in an aprotic solvent (such as DMF) then results in the formation of the desired compound of formula (I). This method is shown in Scheme 1.
The formation of the urethane is typically a two step process but this may also be performed in a one-pot reaction by formation of the activated intermediate in situ.
Scheme 2
(H) (V) (I)
Alternatively, a compound of formula (I) wherein R3 is H can be obtained in a single step by condensation of a (pyridine-4-yl)methanol derivative of formula (II) with the appropriate isocyanate of formula (V) in a suitable solvent (such as DCM), as shown in Scheme 2.
The necessary starting materials for preparing the compounds of formula (I) are either commercially available, or may be prepared by methods known in the art. The processes described below in the experimental section may be carried out to give a compound of the invention in the form of a free base or as an acid addition salt. A pharmaceutically acceptable acid addition salt may be obtained by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds. Examples of addition salt forming acids are mentioned above. - -
The compounds of formula (I) may possess one or more chiral carbon atoms, and they may therefore be obtained in the form of optical isomers, e.g., as a pure enantiomer, or as a mixture of enantiomers (racemate) or as a mixture containing diastereomers. The separation of mixtures of optical isomers to obtain pure enantiomers is well known in the art and may, for example, be achieved by fractional crystallization of salts with optically active (chiral) acids or by chromatographic separation on chiral columns. The chemicals used in the synthetic routes delineated herein may include, for example, solvents, reagents, catalysts, and protecting group and deprotecting group reagents. Examples of protecting groups are t-butoxycarbonyl (Boc), benzyl and trityl (triphenylmethyl). The methods described above may also additionally include steps, either before or after the steps described specifically herein, to add or remove suitable protecting groups in order to ultimately allow synthesis of the compounds. In addition, various synthetic steps may be performed in an alternate sequence or order to give the desired compounds. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing applicable compounds are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser 's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and subsequent editions thereof.
The following abbreviations have been used: aq Aqueous Boc te/t-Butoxy carbonyl
DCM Dichloromethane
DIPEA JV,Λ/-Diisopropylethylamine
DMAP N,Λ/-Dimethylaminopyridine
DMF Λ/,Λ/-Dimethylformamide ES+ Electrospray
Et2O Diethyl ether
EtOAc Ethyl acetate
HIV Human immunodeficiency virus - -
HPLC High performance liquid chromatography
ICV Intracerebroventricular
LCMS Liquid Chromatography Mass Spectrometry
M Molar
[MH]+ Protonated molecular ion
NEt3 Triethylamine
NMM JV-methyl morpholine
RP Reverse Phase sat. Saturated tert Tertiary
TFA Trifluoroacetic acid
THF Tetrahydrofuran
Embodiments of the disclosure are described in the following examples with reference to the accompanying drawings, in which:
Figure 1 is a schematic drawing illustrating weight gain and weight loss in mice during dark and light phases, respectively. The graph illustrates the large nocturnal weight increase versus the comparatively small body weight change over 24 hours
Figure 2 shows the effect of Example 11 on the body weight in mice between the beginning of the dark phase and the beginning of the light phase (pm-am).
Figure 3 shows the effect of Example 15 on the body weight in mice between the beginning of the dark phase and the beginning of the light phase (pm-am).
Figure 4 shows the effect of Example 18 on the body weight in mice between the beginning of the dark phase and the beginning of the light phase (pm-am).
Figure 5 shows the concentration-dependent increase in [3H] -thymidine incorporation by JEG-3 cells for leptin - -
The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.
The disclosure will now be further illustrated by the following non-limiting examples. The specific examples below are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. Without further elaboration, it is believed that one skilled in the art can, based on the description herein, utilize the present disclosure to its fullest extent. All references and publications cited herein are hereby incorporated by reference in their entirety.
- -
EXAMPLES AND INTERMEDIATE COMPOUNDS
Experimental Methods
All reagents were commercial grade and were used as received without further purification, unless otherwise specified. Commercially available anhydrous solvents were used for reactions conducted under inert atmosphere. Reagent grade solvents were used in all other cases, unless otherwise specified. Analytical LCMS was performed on a Waters ZQ mass spectrometer connected to an Agilent 1100 HPLC system. Analytical HPLC was performed on an Agilent 1100 system. High-resolution mass spectra (HRMS) were obtained on an Agilent MSD-TOF connected to an Agilent 1100 HPLC system. During the analyses the calibration was checked by two masses and automatically corrected when needed. Spectra are acquired in positive electrospray mode. The acquired mass range was m/z 100-1100. Profile detection of the mass peaks was used. Normal phase chromatography was performed on a Flash Master Personal system equipped with 2Og Strata SI-I silica gigatubes. Reverse phase chromatography was performed on a Gilson system equipped with Merck LiChoprep® RP- 18 (40-63 μm) 460 x 26mm column, 30 mL/min, gradient of methanol in water from 0% to 100%. Preparative HPLC was performed on a Gilson system equipped with Phenomenex Hydro RP 15O x 20mm, 20 mL/min, gradient of acetonitrile in water from 0% to 100%. The compounds were automatically named using ACD 6.0.
Analytical HPLC and LCMS data were obtained with:
System A: Phenomenex Synergi Hydro RP, (150 x 4.6mm, 4μm), gradient 5-100% CH3CN (+0.085% TFA) in H2O (+0.1% TFA), 1.5 mL/min, gradient time 7 min, 200-300 nm, 30 0C; or
System B: Phenomenex Synergi Hydro RP, (150 x 4.6mm, 4μm), gradient 0-20% CH3CN (+0.085% TFA) in H2O (+0.1% TFA), 1.5 mL/min, gradient time 7 min, 200-300 nm, 30 0C; or System C: Phenomenex Synergi Hydro RP, (150 x 4.6mm, 4μm), gradient 5-100% CH3CN in H2O (+0.1% HCO2H), 1.0 mL/min, gradient time 8 min, 30 °C;or
System D: Phenomenex Synergi Hydro RP, (150 x 4.6mm, 4μm), gradient 0-20% CH3CN in H2O (+0.1% HCO2H), 1.0 mL/min, gradient time 8 min, 30 0C; or - -
System E: Phenomenex Synergi Hydro RP, (150 x 4.6mm, 4μm), gradient 5-100% CH3CN (+0.085% TFA) in H2O (+0.1% TFA), 1.0 niL/min, gradient time 8 min, 30 °C; or System F: Phenomenex Synergi Hydro RP, (30 x 4.6mm, 4μm), gradient 5-100% CH3CN (+0.085% TFA) in H2O (+0.1% TFA), 1.5 niL/min, gradient time 1.75 min, 30 °C.
INTERMEDIATE 1
4-Nitrophenyl (pyridine-4-yl)methyl carbonate
4-Nitrophenyl (pyridine-4-yl)methyl carbonate was prepared according to the procedure described by Veber, D. F., J. Org. Chem. 1977, 42, 3286-3288).
INTERMEDIATE 2
(2,6-Dimethylpyridin-4-yl)methyl 4-nitrophenyl carbonate
A suspension of (2,6-dimethyl-pyridin-4-yl)-methanol (9.14 g, 66.7 mmol; prepared according to the procedure described by Katz, R. B.; Mistry, J.; Mitchell, M. B.; Synthetic Communications, 1989, 19, 317-325) in DCM (40 mL) was added to a solution of Ms-(A- nitrophenyl)carbonate (20.3 g, 66.7 mmol) in DCM (200 mL), followed by NMM (7.34 mL). The reaction mixture was stirred overnight, washed with sat aq NaHCO3 solution (5 x 100 mL), dried (MgSO4) and concentrated in vacuo to give an orange solid. This was recrystallised from EtOAc (~25 mL) to give (2,6-dimethylpyridin-4-yl)methyl A- nitrophenyl carbonate (11.5 g, 57%) as an off white solid. The resulting filtrate was concentrated and the residue obtained was crystallised from EtOAc (15 mL) and heptane (<1 mL) to give (2,6-dimethylpyridin-4-yl)methyl 4-nitrophenyl carbonate (4.76 g, 24%) as an off white solid (81% combined yield). - -
EXAMPLE 1
Pyridin-4-ylmethyl dimethylcarbamate hydrochloride
To a solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 274 mg,
5 1.00 mmol) in DMF (5 mL) was added DIPEA (182 μL, 1.05 mmol), dimethylamine (2M solution in THF, 525 μL, 1.05 mmol) and DMAP (30 mg, catalytic). The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by normal phase chromatography (gradient eluting with MeOH in DCM from 0% to 5%). The residue was dissolved in MeOH (1.0 mL), treated with 2M HCl in Et2O (0.50o mL, 1.0 mmol) and dried in vacuo to give (pyridin-4-yl)methyl dimethylcarbamate hydrochloride (133 mg, 61%) as a white solid.
Analytical HPLC: purity 99.8% (System A, Rτ = 2.85 min); Analytical LCMS: purity 100% (System C, Rτ = 3.43 min), ES+: 180.8 [MH]+; HRMS calcd for C9Hi2N2O2: 180.0899, found 180.0900. 5
EXAMPLE 2
Pyridin-4-ylmethyl [(2S)-tetrahydrofuran-2-ylmethyl] carbamate hydrochloride
To a solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 274 mg,o 1.00 mmol) in DMF (5 mL) was added DIPEA (182 μL, 1.05 mmol), (S)-(tetrahydrofuran- 2-yl)methanamine (108 μL, 1.05 mmol) and DMAP (30 mg, catalytic). The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by normal phase chromatography (gradient eluting with MeOH in DCM, 0 to 5%). The residue was dissolved in MeOH (1.0 mL), treated with 2M HCl in5 Et2O (0.50 mL, 1.0 mmol) and dried in vacuo to give pyridin-4-ylmethyl [(2S)- tetrahydrofuran-2-ylmethyl]carbamate hydrochloride (110 mg, 41%) as an off-white solid. - -
Analytical HPLC: purity 100% (System A, Rτ = 3.07 min); Analytical LCMS: purity 99% (System C, Rτ = 3.56 min), ES+: 236.8 [MH]+; HRMS calcd for Ci2Hi6N2O3: 236.1160, found 236.1166.
EXAMPLE 3
Pyridin-4-ylmethyl (2-hydroxyethyl)carbamate hydrochloride
To a solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 274 mg, 1.0 mmol) in DMF (10 mL) was added DMAP (122 mg, 1.0 mmol) and ethanolamine (62 mg, 1.0 mmol). The reaction mixture was stirred for 48 hours and then concentrated in vacuo. The residue was purified by normal phase chromatography (eluting with EtOAc) and then ion exchange chromatography (1Og Strata-SCX, eluting with MeOH and then 1% NH3 in MeOH). The residue was dissolved in MeOH (1.0 mL), treated with 2M HCl in Et2O (0.50 mL, 1.0 mmol) and dried in vacuo to give pyridin-4-ylmethyl (2-hydroxyethyl)carbamate hydrochloride (24 mg, 10%) as a white powder.
Analytical HPLC: purity 100% (System B, Rτ = 5.48 min); Analytical LCMS: purity 100% (System E, Rτ = 4.24 min), ES+: 196.9 [MH]+; HRMS calcd for C9Hi2N2O3: 196.0848, found 196.0850.
EXAMPLE 4
Pyridin-4-ylmethyl (2-hydroxyethyl)methylcarbamate hydrochloride
To a solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 439 mg, 1.6 mmol) in DMF (10 mL) was added 2-(methylamino)ethanol (150 μL, 1.87 mmol), DIPEA (300 μL, 1.72 mmol) and DMAP (10 mg, catalytic). The reaction mixture was stirred overnight and then concentrated in vacuo. The residue was dissolved in IM aq Na2CO3 solution (25 mL) and extracted with EtOAc (3 x 25 mL). The combined organic phases were dried (MgSO4) and concentrated in vacuo. The residue was purified by normal phase chromatography (gradient eluting with MeOH in DCM from 0% to 10%) and then - -
preparative HPLC (gradient eluting with acetonitrile in water from 0% to 40%). The residue was dissolved in DCM (5 mL), treated with 2M HCl in Et2O (1 mL) and dried in vacuo to give pyridin-4-ylmethyl (2-hydroxyethyl)methylcarbamate hydrochloride (191 mg, 48%) as a white solid.
5 Analytical HPLC: purity 100% (System A, Rτ = 2.55 min); Analytical LCMS: purity 100% (System D, Rτ = 5.40 min), ES+: 211.0 [MH]+; HRMS calcd for Ci0Hi4N2O3: 210.1004, found 210.1007.
EXAMPLE 5 o Pyridin-4-ylmethyl [(lR)-l-(hydroxymethyl)-2-methylpropyl] carbamate
A solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 411 mg, 1.5 mmol), (i?)-(-)-2-amino-3-methyl-l-butanol (155 mg, 1.5 mmol) and DMAP (183 mg, 1.5 mmol) in DMF (10 mL) were stirred overnight. The DMF was removed in vacuo. Thes residue was taken up in EtOAc and washed with water (x 2), aq Na2CO3 solution (x 4), dried (MgSO4) and concentrated in vacuo. The desired product was crystallised from Et2O, further purified by reverse phase chromatography and then dried in vacuo to give pyridin- 4-ylmethyl [(I R)- l-(hydroxymethyl)-2-methylpropyl] carbamate (70 mg, 20%) as a white crystalline solid. o Analytical HPLC: purity 100% (System A, Rτ = 3.13 min); Analytical LCMS: purity 99% (System C, Rτ = 3.71 min), ES+: 239.0 [MH]+.
EXAMPLE 6
Pyridin-4-ylmethyl [(lR)-l-(hydroxymethyl)-3-methylbutyl] carbamate
To a solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 411 mg, 1.5 mmol) and (i?)-leucinol (176 mg, 1.5 mmol) in DMF (5 mL) was added DMAP (183 mg, 1.5 mmol) and left to stir for 24 hours. The DMF was removed in vacuo. The residue was - -
taken up in EtOAc and washed with water, aq Na2CO3 solution (x 8), dried (MgSO4) and concentrated in vacuo. The residue was purified by normal phase chromatography (1O g RediSep column, 20 mL/min, gradient eluting with MeOH in DCM from 0% to 5%) and dried in vacuo to give pyridin-4-ylmethyl [(lR)-l-(hydroxymethyl)-3- methylbutyljcarbamate (75 mg, 20%) as a white crystalline solid.
Analytical HPLC: purity 99% (System A, Rτ = 3.54 min); Analytical LCMS: purity >99% (System C, Rτ = 4.07 min), ES+: 253.0 [MH]+; HRMS calcd for Ci3H20N2O3: 252.1474, found 252.1481.
EXAMPLE 7
Pyridin-4-ylmethyl cyclopentylcarbamate
To a solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 2.74 g, 10 mmol) in DMF (30 mL) was added DIPEA (1.75 mL, 10 mmol), cyclopentylamine (0.99 mL, 10 mmol) followed by DMAP (50 mg, catalytic). The reaction mixture was stirred at room temperature overnight and then dried in vacuo. The residue was dissolved in EtOAc (150 mL) and washed with sat aq KHCO3 solution (6 x 150 mL), brine (100 mL), dried (MgSO4) and concentrated in vacuo. The residue was purified by normal phase chromatography (40 g Biotage, gradient eluting with MeOH in DCM from 0% to 5%). The impure fractions were combined and re-purified by normal phase chromatography (gradient eluting with MeOH in DCM from 0% to 5%). The pure fractions from both columns were combined, crystallised from ether and dried in vacuo to give pyridin-4- ylmethyl cyclopentylcarbamate (1.08 g, 49%) as a white solid. Analytical HPLC: purity 100% (System A, Rτ = 3.73 min); Analytical LCMS: purity 99.3% (System C, Rτ = 4.44 min), ES+: 220.9 [MH]+; HRMS calcd for Ci2Hi6N2O2: 220.1212, found 220.1214. - -
EXAMPLE 8
Pyridin-4-ylmethyl (3R)-tetrahydrofuran-3-ylcarbamate
To a solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 274 mg, 1.0 mmol) and DMAP (122 mg, 1.0 mmol) in DMF (5 mL) was added a solution of (R)- tetrahydrofuran-3 -amine toluenesulfonate (259 mg, 1.0 mmol) and DIPEA (174 μL, 1.0 mmol) in DMF (5 mL). The reaction mixture was stirred for 24 hours and then concentrated in vacuo. The residue was purified by normal phase chromatography
(gradient eluting with MeOH in DCM from 0% to 2%) and dried in vacuo to give pyridin- 4-ylmethyl (3R)-tetrahydrofuran-3-ylcarbamate (68 mg, 31%) as an off white solid.
Analytical HPLC: purity >99% (System A, Rτ = 2.72 min); Analytical LCMS: purity 100% (System C, Rτ = 3.32 min), ES+: 222.8 [MH]+; HRMS calcd for CnHi4N2O3: 222.1004, found 222.1007.
EXAMPLE 9
Pyridin-4-ylmethyl [(l-hydroxycyclohexyljmethyl] carbamate
To a solution of 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 274 mg, 1.0 mmol) and DMAP (122 mg, 1.0 mmol) in DMF (10 mL) was added a solution of DIPEA (175 μL, 1.0 mmol) and 1-aminomethylcyclohexanol hydrochloride (166 mg, 1.0 mmol) in DMF (10 mL). The reaction mixture was stirred for 18 hours and then concentrated in vacuo. The residue was taken up in EtOAc and washed with IM aq Na2CO3 solution to remove p-nitrophenol, dried (Na2SO4) and concentrated in vacuo. The residue was purified by normal phase chromatography (eluting with EtOAc) and dried in vacuo to give pyridin- 4-ylmethyl [(l-hydroxycyclohexyl)methyl]carbamate (195 mg, 74%) as a light brown oil. Analytical HPLC: purity 100% (System A, Rτ = 3.54 min); Analytical LCMS: purity 99% (System C, Rτ = 4.18 min), ES+: 265.0 [MH]+; HRMS calcd for Ci4H20N2O3: 264.1474, found 264.1479. - -
EXAMPLE 10
(Pyridin-4-yl)methyl (lR,2S)-2,3-dihydro-2-hydroxy-lH-inden-l-ylcarbamate hydrochloride
To a solution of (li?,2S)-(+)-cώ-l-amino-2-indanol (149 mg, 1.0 mmol), 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 274 g, 1.0 mmol) and DIPEA (354 μL, 2.0 mmol) in DMF (5 mL) was added DMAP (catalytic amount). The reaction mixture was stirred at room temperature for 16 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (50 mL) and washed with IM aq Na2CO3 solution (5 x 30 mL), dried (MgSO4) and concentrated in vacuo. The residue was purified by normal phase chromatography (gradient eluting with MeOH in DCM from 0% to 3%). The residue was dissolved in DCM (30 mL) and excess 2M ΗCI in Et2O was added. The resulting precipitate was collected by filtration and dried in vacuo to give (pyridin-4-yl)methyl (li?,25)-2,3-dihydro-2-hydroxy-l/f-inden-l-ylcarbamate hydrochloride (70 mg, 22%) as a white solid.
Analytical ΗPLC: purity 97.1% (System A, Rτ = 5.61 min); Analytical LCMS: purity 100% (System C, Rτ = 4.07 min), ES+: 285.0 [MH]+. HRMS calcd for Ci6Hi6N2O3: 284.1161, found 284.1164.
EXAMPLE 11
Pyridin-4-ylmethyl phenylcarbamate
To a stirred solution of (pyridin-4-yl)methanol (545 mg, 5.0 mmol) in DCM (10 mL) was added phenyl isocyanate (596 mg, 5.0 mmol). The reaction mixture was stirred overnight, washed with water (20 mL) and then concentrated in vacuo. The residue was purified by normal phase chromatography (250x25 mm column packed with ICN silica (18-32, 6θA), 30 mL/min, gradient eluting with MeOH in DCM from 0 to 5%). The residue was taken up in Et2O and a white crystalline solid formed. The crystals were collected by filtration and - -
dried in vacuo to give pyridin-4-ylmethyl phenylcarbamate (477 mg, 42%) as a white crystalline solid.
Analytical HPLC: purity 100% (System A, Rτ = 3.95 min); Analytical LCMS: purity 100% (System E, Rτ = 5.70 min), ES+: 229.3 [MH]+; HRMS calcd for Ci3Hi2N2O2: 228.0899, found 228.0901
EXAMPLE 12
Pyridin-4-ylmethyl [(lS)-l-phenylethyl]carbamate hydrochloride
HCl 4-Nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 274 mg, 1.0 mmol), (S)-I- methylbenzylamine (122 mg, 1.0 mmol) and DMAP (122 mg, 1.0 mmol) were dissolved in DMF (10 mL). The reaction mixture was stirred for 20 hours and then concentrated in vacuo. The residue was taken up in EtOAc and washed with a IM aq Na2CO3 solution to remove p-nitrophenol, dried (MgSO4) and concentrated in vacuo. The residue was purified by normal phase chromatography (eluting with EtOAc) and then reverse phase chromatography. The residue was dissolved in DCM (4 mL), treated with 2M HCl in Et2O (2.0 mL, 4.0 mmol) and dried in vacuo to give pyridin-4-ylmethyl [(I S)-I- phenylethyljcarbamate hydrochloride (136 mg, 46%) as a white solid. Analytical HPLC: purity 100% (System A, Rτ = 4.11 min); Analytical LCMS: purity 100% (System C, Rτ = 4.68 min), ES+: 257.0 [MH]+; HRMS calcd for Ci5Hi6N2O2: 256.1212, found 256.1216.
EXAMPLE 13
Pyridin-4-ylmethyl [(lR)-2-hydroxy-l-phenylethyl] carbamate
To a solution of (i?)-phenylglycinol (1.54 g, 11.2 mmol) in DMF (75 mL) was added DIPEA (2.6 mL, 15.0 mmol), 4-nitrophenyl (pyridin-4-yl)methyl carbonate (Intermediate 1; 2.98 g, 10.9 mmol) and then DMAP (60 mg, catalytic). The reaction mixture was stirred - -
for 18 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (150 mL), washed with IM aq Na2CO3 solution (5 x 100 mL), dried (MgSO4) and concentrated in vacuo. The residue was purified by normal phase chromatography (gradient eluting with MeOH in DCM from 0% to 5%) and dried in vacuo to give pyridin-4-ylmethyl [(lR)-2- hydroxy- l-phenylethyl]carbamate (1.92 g, 68%) as a white powder.
Analytical HPLC: purity 99.4% (System A, Rτ = 3.47 min); Analytical LCMS: purity 99% (System C, Rτ = 3.97 min), ES+: 272.7 [MH]+; HRMS calcd for Ci5Hi6N2O3: 272.1161, found 272.1168.
EXAMPLE 14
(2,6-Dimethylpyridin-4-yl)methyl (l-methyl-l-phenylethyl)carbamate hydrochloride
To a solution of (2,6-dimethylpyridin-4-yl)methyl 4-nitrophenyl carbonate (Intermediate 2; 615 mg, 2.0 mmol) in DMF (6 mL) was added DIPEA (350 μL, 2.0 mmol), 1,1- dimethylbenzylamine (277 mg, 2.0 mmol) and then DMAP (10 mg, catalytic). The reaction mixture was stirred overnight and then concentrated in vacuo. The residue was purified by normal phase chromatography (gradient eluting with MeOH in DCM from 0% to 10%) followed by reverse phase chromatography. The residue was dissolved in Et2O (5 mL), treated with 2M HCl in Et2O (1.0 mL, 2.0 mmol) and dried in vacuo to give (2,6- dimethylpyridin-4-yl)methyl (1 -methyl- l-phenylethyl)carbamate hydrochloride (284 mg, 42%) as a white powder.
Analytical HPLC: purity 100% (System A, Rτ = 4.64 min); Analytical LCMS: purity 100% (System C, Rτ = 5.52 min), ES+: 299.4 [MH]+.
EXAMPLE 15
(2,6-Dimethylpyridin-4-yl)methyl ført-butylcarbamate hydrochloride
- -
(2,6-Dimethylpyridin-4-yl)methyl 4-nitrophenyl carbonate (Intermediate 2; 616 mg, 2.0 mmol), tert-butylamine (500 μL, 4.8 mmol), DIPEA (400 μL, 2.3 mmol) and DMAP (10 mg, catalytic) were dissolved in DMF (5 mL) and stirred overnight. The reaction mixture was concentrated in vacuo and the residue dissolved in EtOAc (30 mL), washed with IM
5 aq Na2CCh solution (4 x 30 mL), dried (MgSO4) and concentrated in vacuo. The crude product was purified by reverse phase chromatography (gradient eluting with MeOH in water from 0% to 100%, with 1% formic acid in each solvent). The residue was dissolved in DCM (5 mL), treated with 2M HCl in Et2O (1.0 mL, 2.0 mmol) and dried in vacuo to give (2,6-dimethylpyridin-4-yl)methyl tert-butylcarbamate hydrochloride as a whiteo powder (298 mg, 54%).
Analytical HPLC: purity 100% (System A, Rτ = 4.08 min); Analytical LCMS: purity 100% (System C, Rτ = 4.33 min), ES+: 237.2 [MH]+; HRMS calcd for Ci3H20N2O2: 236.1525, found 236.1533. s EXAMPLE 16
(2,6-Dimethylpyridin-4-yl)methyl cyclopentylcarbamate hydrochloride
To a stirred solution of (2,6-dimethylpyridin-4-yl)methyl 4-nitrophenyl carbonate (Intermediate 2; 537 mg, 1.78 mmol) in DMF (6 mL) was added DIPEA (320 μL, 1.84o mmol), cyclopentylamine (175 μL, 1.78 mmol) and then DMAP (10 mg, catalytic). The reaction mixture was stirred for five days and then concentrated in vacuo. The residue was purified by normal phase chromatography (gradient eluting with MeOH in DCM from 0% to 10%) followed by reverse phase chromatography. The residue was dissolved in Et2O (5 mL), treated with 2M HCl in Et2O (1.0 mL, 2.0 mmol) and dried in vacuo to give (2,6-5 dimethylpyridin-4-yl)methyl cyclopentylcarbamate hydrochloride (280 mg, 55%) as a white solid.
Analytical HPLC: purity 100% (System A, Rτ = 4.10 min); Analytical LCMS: purity 100% (System C, Rτ = 5.05 min), ES+: 249.2 [MH]+; HRMS calcd for Ci4H20N2O2: 248.1525, found 248.1533. 0 - -
EXAMPLE 17
(2,6-Dimethylpyridin-4-yl)methyl (cyclopropylmethyljcarbamate hydrochloride
To a solution of (2,6-dimethylpyridin-4-yl)methyl 4-nitrophenyl carbonate (Intermediate 2; 575 mg, 1.9 mmol) in DMF (5 mL) was added cyclopropylmethanamine (440 μL, 4.7 mmol), DIPEA (400 μL, 2.3 mmol) and DMAP (10 mg, catalytic). The reaction mixture was stirred for 3 days and then concentrated in vacuo. The residue was dissolved in EtOAc (25 mL), washed with IM aq Na2CO3 solution (4 x 25 mL), dried (MgSO4) and concentrated in vacuo. The residue was purified by reverse phase chromatography (gradient eluting with MeOH in water from 0% to 100%, with 1% formic acid in each solvent). The residue was dissolved in DCM (5 mL), treated with 2M HCl in Et2O (1.0 mL, 2.0 mmol) and dried in vacuo to give (2,6-dimethylpyridin-4-yl)methyl (cyclopropylmethyl)carbamate hydrochloride (302 mg, 59%) as a white powder. Analytical HPLC: purity 100% (System A, Rτ = 3.77 min); Analytical LCMS: purity 99.4% (System C, Rτ = 4.13 min), ES+: 235.5 [MH]+; HRMS calcd for Ci3Hi8N2O2: 234.1368, found 234.1371.
EXAMPLE 18
(2,6-Dimethylpyridin-4-yl)methyl (3R)-tetrahydrofuran-3-ylcarbamate hydrochloride
To a solution of (2,6-dimethylpyridin-4-yl)methyl 4-nitrophenyl carbonate (Intermediate 2;
235 mg, 0.78 mmol) in DMF (5 mL) was added DIPEA (400 μL, 2.3 mmol), (R)- tetrahydrofuran-3 -amine toluenesulfonate (221 mg, 0.85 mmol) and DMAP (10 mg, catalytic). The reaction mixture was stirred overnight and then concentrated in vacuo. The residue was dissolved in EtOAc (25 mL), washed with IM aq Na2CO3 solution (5 x 25 mL), dried (MgSO4) and concentrated in vacuo. The residue was dissolved in DCM (5 mL) and treated with 2M HCl in Et2O (0.5 mL, 1.0 mmol) to give a precipitate. The - -
solvent was removed by decantation and the precipitate was dried in vacuo to give (2,6- dimethylpyridin-4-yl)methyl (3R)-tetrahydrofuran-3-ylcarbamate hydrochloride (102 mg, 46%) as a white solid.
Analytical HPLC: purity 99.3% (System A, Rτ = 3.03 min); Analytical LCMS: purity 100% (System E, Rτ = 4.59 min), ES+: 251.4 [MH]+; HRMS calcd for Ci3Hi8N2O3: 250.1317, found 250.1330.
EXAMPLE 19
(2,6-Dimethylpyridin-4-yl)methyl (3,5-dimethylisoxazol-4-yl)carbamate hydrochloride
To a stirred solution of (2,6-dimethylpyridin-4-yl)methanol (274 mg, 2.0 mmol; prepared according to the procedure described by Katz, R. B.; Mistry, J.; Mitchell, M. B.; Synthetic Communications, 1989, 19, 317-325) in DCM (10 mL) was added 4-isocyanato-3,5- dimethylisoxazole (276 mg, 2.0 mmol). After stirring overnight at room temperature the reaction mixture was concentrated in vacuo. The residue was purified by normal phase chromatography (gradient eluting with MeOH in DCM from 0% to 5%). The residue was dissolved in Et2O, treated with 2M HCl in dioxane (1.0 mL, 2.0 mmol) and then concentrated in vacuo. The residue was taken up in water and dried in vacuo to give (2,6- dimethylpyridin-4-yl)methyl (3,5-dimethylisoxazol-4-yl)carbamate hydrochloride (250 mg, 40%) as a white crystalline solid.
Analytical HPLC: purity 100% (System A, Rτ = 3.35 min); Analytical LCMS: purity 100% (System E, Rτ = 5.07 min), ES+: 276.4 [MH]+; HRMS calcd for Ci4Hi7N3O3: 275.1270, found 275.1279. - -
BIOLOGICAL TESTS
Measurement of overnight body weight change in male C57 bl/6 mice
This model studies the effects of compounds on body weight gain during the pm-am period in order to maximise the effective window. Typically the mice gain about 1 g in weight during the dark phase and then loose the majority of this weight gain during the light phase, as represented in Figure 1. The weight difference over any 24 hour period is very small whilst the weight difference between the beginning of the dark phase and the beginning of the light phase (pm-am) is maximal.
It is important to measure body weight change over the dark phase. If mice are dosed with an active compound on two consecutive days and the bodyweight change is recorded 48 hours after the first dose then no significant effect is observed. However if the body weight change over the dark phase only is considered a significant and robust effect is seen. This is because the mice rebound during the light phase to compensate for the lack of weight gain over the dark phase. Very active long lasting compounds may also diminish this rebound and reduce the body weight over the 48 hours.
Weight change over consecutive days in C57bl\6 male mice:
The weight difference between the beginning of the dark phase and the beginning of the light phase (pm-am) is greater than the weight difference measured between pm and pm on 2 consecutive days. The effect of the compounds on the pm-am difference was therefore studied in order to maximise the effect window.
C57 bl/6 mice were grouped (5 per cage) and left 5 days for acclimatisation. A single intraperitoneally (ip) administered dose (60 mg/kg) was given just prior to the dark phase. Compounds were either water soluble or dissolved in up to 3% cremophor (in this case the vehicle also contained cremophor). The pH was adjusted from a minimum of 5.5 to a maximum of 8 depending on the nature of the compound.
As shown in Figures 2-4, compounds of Formula (I) are useful for decreasing body weight in mice. - -
Leptin assay in non-recombinant system
Although well-characterised in recombinant systems (e.g. ObRb-transfected HEK293 cells), where leptin elicits a very marked increase in STAT3 phosphorylation, these systems have often failed to provide an accurate measure of activity of a test compound towards the leptin receptor. It seems that overexpression of the receptor (as well as the possibility for different drugs to act on different parts of the signaling pathway triggered by leptin association with its receptor) results in most cases in the absence of activity of the drugs tested.
The leptin receptor expression in non-recombinant system is often fluctuating and care must be given to identify a system where signal stability remains within experiments. Using such a system, leptin receptor antagonist mimetics could be identified by evaluating their action vs. leptin (see below).
Leptin is produced chiefly in adipose cells, but in humans, mRNA encoding leptin is also present in the placenta. Here, leptin might play an important proliferative role in the microvasculature. The possibility to use this hypothesis in a native cell line was evaluated.
JEG-3 protocol
In JEG-3 cells (choriocarcinoma cell line) leptin is able to stimulate proliferation up to 3 fold (Biol. Reprod. (2007) 76: 203-10). Leptin also causes a concentration-dependent increase in [ H]-thymidine incorporation in JEG-3 cells (Figure 5, maximal effect at 100 nM (EC50 = 2.1 nM)). The radioactivity incorporated by the cells is an index of their proliferative activity and is measured in counts per minute (CPM) with a liquid scintillation beta counter. This finding can be applied to test whether a compound is able to either reproduce the effect of leptin on cell proliferation (leptin receptor agonist mimetic) (i.e., a given compound will cause an increase in incorporated [3H] -Thymidine by the cells) or to inhibit the effect of leptin (antagonistic effect) by preventing the leptin-mediated increase in [ H]- thymidine incorporation. - -
This approach has the advantage of using a non-recombinant system and has reasonable reproducibility and robustness.
Measurement of brain penetration
The test species (rodent) is given a bolus dose of the substrate under investigation, usually via intravenous (IV) or oral (PO) routes. At appropriate time points, blood samples are taken and the resultant plasma extracted and analysed for substrate concentration and, where appropriate, metabolite concentration. At similar time points, animals from another group are sacrificed, brains isolated and the brain surface cleaned. Brain samples are then homogenised, extracted and analysed for substrate concentration and, where appropriate, metabolite concentration. Alternatively, microdialysis probes are implanted into one or more brain regions of the test species and samples collected at appropriate time points for subsequent analysis. This method has the advantage of measuring only extra-cellular substrate concentration. Plasma and brain concentrations are then compared and ratios calculated, either by comparison of averaged concentrations at individual time points, or by calculation of the area-under-the-curve (AUC) of the concentration-time plots.

Claims

- -CLAIMS
1. A compound of formula (I)
or a pharmaceutically acceptable salt, solvate, hydrate, geometrical isomer, tautomer or optical isomer thereof, wherein:
each R1 is independently selected from Ci_4-alkyl, Ci_4-alkoxy, halogen, cyano and
CF3;
R2 is Ci_6-alkyl (optionally substituted with one or more substituents selected from hydroxy, halogen and cyano) or -[C(R4A)(R4B)]m-R5;
R3 is hydrogen, Ci_4-alkyl or fluoro-Ci_4-alkyl; R4A and R4B are each independently selected from hydrogen, halogen, hydroxy, C1-4- alkyl, fluoro-Ci_4-alkyl and hydroxy-Ci_4-alkyl;
R5 is C3-8-cycloalkyl, Cβ-io-aryl, heterocyclyl or heteroaryl, each of which is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro, CF3 and Ci_4-alkyl;
m is 0, 1 or 2; and n is O, 1, 2, 3 or 4;
with the proviso that the compound is not selected from: • pyridin-4-ylmethyl bis(2-chloroethyl)carbamate;
• (2,6-dichloropyridin-4-yl)methyl dimethylcarbamate;
• (2,6-dichloropyridin-4-ylmethyl propylcarbamate;
• pyridin-4-ylmethyl methylcarbamate;
• pyridin-4-ylmethyl isopropylcarbamate; • pyridin-4-ylmethyl [3 -(trifluoromethyl)phenyl] carbamate; - -
• pyridin-4-ylmethyl (5-chloro-2-methoxyphenyl)carbamate;
• pyridin-4-ylmethyl (2-methoxyphenyl)carbamate;
• pyridin-4-ylmethyl (2,6-dimethylphenyl)carbamate;
• pyridin-4-ylmethyl (2,4,6-trimethylphenyl)carbamate; • pyridin-4-ylmethyl (5-chloro-2,4-dimethoxyphenyl)carbamate;
• pyridin-4-ylmethyl (2-methyl-5-nitrophenyl)carbamate;
• pyridin-4-ylmethyl (3-ethylphenyl)carbamate;
• pyridin-4-ylmethyl (2,4-dimethylphenyl)carbamate;
• pyridin-4-ylmethyl (3,4,5-trimethoxyphenyl)carbamate; • pyridin-4-ylmethyl cyclohexyl(methyl)carbamate;
• pyridin-4-ylmethyl pyridin-3-ylcarbamate;
• pyridin-4-ylmethyl (4-methoxyphenyl)carbamate;
• pyridin-4-ylmethyl (2,6-dichlorophenyl)carbamate;
• pyridin-4-ylmethyl cyclohexylcarbamate; • pyridin-4-ylmethyl pyridin-4-ylcarbamate;
• pyridin-4-ylmethyl (4-fluorophenyl)carbamate;
• pyridin-4-ylmethyl [(2-chlorophenyl)methyl] carbamate;
• pyridin-4-ylmethyl (3-nitrophenyl)carbamate;
• pyridin-4-ylmethyl (3,5-dimethoxyphenyl)carbamate; • pyridin-4-ylmethyl (4-methylphenyl)carbamate;
• pyridin-4-ylmethyl (3-chlorophenyl)carbamate;
• pyridin-4-ylmethyl (4-chlorophenyl)carbamate;
• (2,6-dichloropyridin-4-yl)methyl phenylcarbamate;
• (2,6-dimethylpyridin-4-yl)methyl phenylcarbamate; • pyridin-4-ylmethyl phenylcarbamate; and
• pyridin-4-ylmethyl (3,4-dimethoxyphenyl)carbamate.
2. A compound according to claim 1, wherein R2 is optionally substituted Ci_6-alkyl.
3. A compound according to claim 1, wherein R2 is -[C(R4A)(R4B)]m-R5 and wherein m is 0 or 1.
4. A compound according to claim 1, which is selected from: - -
• pyridin-4-ylmethyl dimethylcarbamate;
• pyridin-4-ylmethyl [(2S)-tetrahydrofuran-2-ylmethyl]carbamate;
• pyridin-4-ylmethyl (2-hydroxyethyl)carbamate;
• pyridin-4-ylmethyl (2-hydroxyethyl)methylcarbamate; • pyridin-4-ylmethyl [(li?)-l-(hydroxymethyl)-2-methylpropyl]carbamate;
• pyridin-4-ylmethyl [(IR)- 1 -(hydroxymethyl)-3-methylbutyl]carbamate;
• pyridin-4-ylmethyl cyclopentylcarbamate;
• pyridin-4-ylmethyl (3i?)-tetrahydrofuran-3-ylcarbamate;
• pyridin-4-ylmethyl [(l-hydroxycyclohexy^methyljcarbamate; • (pyridin-4-yl)methyl (li?,2S)-2,3-dihydro-2-hydroxy-lH-inden-l-ylcarbamate;
• pyridin-4-ylmethyl [( 1 S)- 1 -phenylethyl] carbamate;
• pyridin-4-ylmethyl [(li?)-2-hydroxy-l-phenylethyl]carbamate;
• (2,6-dimethylpyridin-4-yl)methyl (1 -methyl- 1 -phenylethyl)carbamate;
• (2,6-dimethylpyridin-4-yl)methyl tert-butylcarbamate; • (2,6-dimethylpyridin-4-yl)methyl cyclopentylcarbamate;
• (2,6-dimethylpyridin-4-yl)methyl (cyclopropylmethyl)carbamate;
• (2,6-dimethylpyridin-4-yl)methyl (3i?)-tetrahydrofuran-3-ylcarbamate; and
• (2,6-dimethylpyridin-4-yl)methyl (3,5-dimethylisoxazol-4-yl)carbamate.
5. A pharmaceutical formulation containing a compound according to any one of claims 1 to 4 as active ingredient, in combination with a pharmaceutically acceptable diluent or carrier.
6. A compound according to any one of claims 1 to 4 for use in therapy.
7. A compound of formula (I)
- -
or a pharmaceutically acceptable salt, solvate, hydrate, geometrical isomer, tautomer or optical isomer thereof, wherein:
each R1 is independently selected from Ci_4-alkyl, Ci_4-alkoxy, halogen, cyano and CF3;
R2 is Ci_6-alkyl (optionally substituted with one or more substituents selected from hydroxy, halogen and cyano) or -[C(R4A)(R4B)]m-R5;
R3 is hydrogen, Ci_4-alkyl or fluoro-Ci_4-alkyl;
R4A and R4B are each independently selected from hydrogen, halogen, hydroxy, C1-4- alkyl, fluoro-Ci_4-alkyl and hydroxy-Ci_4-alkyl;
R5 is C3-8-cycloalkyl, Cβ-io-aryl, heterocyclyl or heteroaryl, each of which is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro, CF3 and Ci_4-alkyl; m is 0, 1 or 2; and n is 0, 1, 2, 3 or 4,
for use in the treatment or prevention of conditions or diseases that are prevented, treated, or ameliorated by selective action via the leptin receptor.
8. A compound according to claim 7, for use in the treatment or prevention of conditions or diseases associated with weight gain.
9. The compound according to claim 8, wherein the condition or disease is obesity, type 2 diabetes, lipodystrophy, insulin resistance, metabolic syndrome, hyperglycemia, hyperinsulinemia, dyslipidemia, hepatic steatosis, hyperphagia, hypertension, hypertriglyceridemia, infertility, a skin disorder associated with weight gain or macular degeneration.
10. A compound according to claim 7 for use in the treatment or prevention of severe weight loss, dysmenorrhea, amenorrhea, female infertility or immunodeficiency, or in the treatment of wound healing. - -
11. A compound according to claim 7 for use in the treatment or prevention of inflammatory conditions or diseases, low level inflammation associated with obesity and excess plasma leptin, atherosclerosis, macro or micro vascular complications of type 1 or 2 diabetes, retinopathy, nephropathy, autonomic neuropathy, or blood vessel damage caused by ischaemia or atherosclerosis.
12. A compound according to claim 7 for use in the inhibition of angiogenesis.
13. Use of a compound of formula (I)
or a pharmaceutically acceptable salt, solvate, hydrate, geometrical isomer, tautomer or optical isomer thereof, wherein:
each R1 is independently selected from Ci_4-alkyl, Ci_4-alkoxy, halogen, cyano and
CF3;
R2 is Ci_6-alkyl (optionally substituted with one or more substituents selected from hydroxy, halogen and cyano) or -[C(R4A)(R4B)]m-R5; R3 is hydrogen, Ci_4-alkyl or fluoro-Ci_4-alkyl;
R4A and R4B are each independently selected from hydrogen, halogen, hydroxy, C1-4- alkyl, fluoro-Ci_4-alkyl and hydroxy-Ci_4-alkyl;
R5 is C3-8-cycloalkyl, Cβ-io-aryl, heterocyclyl or heteroaryl, each of which is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro, CF3 and Ci_4-alkyl; m is 0, 1 or 2; and n is 0, 1, 2, 3 or 4, - -
in the manufacture of a medicament for the treatment or prevention of conditions or diseases that are prevented, treated, or ameliorated by selective action via the leptin receptor.
14. The use according to claim 13, for the treatment or prevention of conditions or diseases associated with weight gain.
15. The use according to claim 14, wherein the condition or disease is obesity, type 2 diabetes, lipodystrophy, insulin resistance, metabolic syndrome, hyperglycemia, hyperinsulinemia, dyslipidemia, hepatic steatosis, hyperphagia, hypertension, hypertriglyceridemia, infertility, a skin disorder associated with weight gain or macular degeneration.
16. The use according to claim 13, for the treatment or prevention of severe weight loss, dysmenorrhea, amenorrhea, female infertility or immunodeficiency, or for the treatment of wound healing.
17. The use according to claim 13, for the treatment or prevention of inflammatory conditions or diseases, low level inflammation associated with obesity and excess plasma leptin, atherosclerosis, macro or micro vascular complications of type 1 or 2 diabetes, retinopathy, nephropathy, autonomic neuropathy, or blood vessel damage caused by ischaemia or atherosclerosis.
18. The use according to claim 13, for the inhibition of angiogenesis.
19. A method for the treatment or prevention of conditions or diseases that are prevented, treated, or ameliorated by selective action via the leptin receptor, which comprises administering to a mammal, including man, in need of such treatment an effective amount of a compound of formula (I) - -
or a pharmaceutically acceptable salt, solvate, hydrate, geometrical isomer, tautomer or optical isomer thereof, wherein:
each R1 is independently selected from Ci_4-alkyl, Ci_4-alkoxy, halogen, cyano and
CF3;
R2 is Ci_6-alkyl (optionally substituted with one or more substituents selected from hydroxy, halogen and cyano) or -[C(R4A)(R4B)]m-R5; R3 is hydrogen, Ci_4-alkyl or fluoro-Ci_4-alkyl;
R4A and R4B are each independently selected from hydrogen, halogen, hydroxy, C1-4- alkyl, fluoro-Ci_4-alkyl and hydroxy-Ci_4-alkyl;
R5 is C3_8-cycloalkyl, Cβ-io-aryl, heterocyclyl or heteroaryl, each of which is optionally substituted with one or more substituents selected from hydroxy, halogen, cyano, nitro, CF3 and Ci_4-alkyl; m is 0, 1 or 2; and n is 0, 1, 2, 3 or 4.
20. The method according to claim 19, for the treatment or prevention of conditions or diseases associated with weight gain.
21. The method according to claim 20, wherein the condition or disease is obesity, type 2 diabetes, lipodystrophy, insulin resistance, metabolic syndrome, hyperglycemia, hyperinsulinemia, dyslipidemia, hepatic steatosis, hyperphagia, hypertension, hypertriglyceridemia, infertility, a skin disorder associated with weight gain or macular degeneration.
22. The method according to claim 19, for the treatment or prevention of severe weight loss, dysmenorrhea, amenorrhea, female infertility or immunodeficiency, or for the treatment of wound healing. - -
23. The method according to claim 19, for the treatment or prevention of inflammatory conditions or diseases, low level inflammation associated with obesity and excess plasma leptin, atherosclerosis, macro or micro vascular complications of type 1 or 2 diabetes, retinopathy, nephropathy, autonomic neuropathy, or blood vessel damage caused by ischaemia or atherosclerosis.
24. The method according to claim 19, for inhibition of angiogenesis.
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